Aminated psilocybin derivatives and methods of using

ABSTRACT

Disclosed are novel aminated psilocybin derivative compounds and pharmaceutical and recreational drug formulations containing the same. The aminated psilocybin derivative compounds may be chemically synthesized.

RELATED APPLICATION

This application is a continuation of PCT Application No.PCT/CA2022/050007 filed Jan. 5, 2022, which claims the benefit of U.S.Provisional Application No. 63/248,009 filed Sep. 24, 2021; the entirecontents of Patent Application Nos. PCT/CA2022/050007 and 63/248,009 arehereby incorporated by reference.

INCORPORATION OF SEQUENCE LISTING

A computer readable form of the Sequence Listing“29664-P62652US03_SequenceListing.xml” (29,876 bytes), submitted viaEFS-WEB and created on Sep. 15, 2022, is herein incorporated byreference.

FIELD OF THE DISCLOSURE

The compositions and methods disclosed herein relate to a chemicalcompound known as psilocybin. Furthermore, the compositions and methodsdisclosed herein relate in particular to aminated forms of psilocybin.

BACKGROUND OF THE DISCLOSURE

The following paragraphs are provided by way of background to thepresent disclosure. They are not however an admission that anythingdiscussed therein is prior art or part of the knowledge of a person ofskill in the art.

The biochemical pathways in the cells of living organisms may beclassified as being part of primary metabolism, or as being part ofsecondary metabolism. Pathways that are part of a cell's primarymetabolism are involved in catabolism for energy production or inanabolism for building block production for the cell. Secondarymetabolites, on the other hand, are produced by the cell without havingan obvious anabolic or catabolic function. It has long been recognizedthat secondary metabolites can be useful in many respects, including astherapeutic compounds.

Psilocybin, for example, is a secondary metabolite that is naturallyproduced by certain mushrooms which taxonomically can be classified asbelonging the Basidiomycota division of the fungi kingdom. Mushroomspecies which can produce psilocybin include species belonging to thegenus Psilocybe, such as Psilocybe azurescens, Psilocybe semilanceata,Psilocybe serbica, Psilocybe mexicana, and Psilocybe cyanescens, forexample. The interest of the art in psilocybin is well established.Thus, for example, psilocybin is a psychoactive compound and istherefore used as a recreational drug. Furthermore, psilocybin is usedas a research tool in behavioral and neuro-imaging studies in psychoticdisorders, and has been evaluated for its clinical potential in thetreatment of mental health conditions (Daniel, J. et al. Mental HealthClin/, 2017; 7(1): 24-28), including to treat anxiety in terminal cancerpatients (Grob, C. et al. Arch. Gen. Psychiatry, 2011, 68(1) 71-78) andto alleviate symptoms of treatment-resistant depression (Cathart-Harris,R. L. et al. Lancet Psychiatry, 2016, 3: 619-627).

Although the toxicity of psilocybin is low, adverse side effects,including, for example, panic attacks, paranoia, and psychotic states,sometimes together or individually referred to as “a bad trip”, are notinfrequently experienced by recreational psilocybin users.

There exists therefore a need in the art for improved psilocybincompounds.

SUMMARY OF THE DISCLOSURE

The following paragraphs are intended to introduce the reader to themore detailed description, not to define or limit the claimed subjectmatter of the present disclosure.

In one aspect, the present disclosure relates to psilocybin andderivative compounds.

In another aspect, the present disclosure relates to aminated psilocybinderivative compounds and methods of making and using these compounds.

Accordingly, in one aspect, the present disclosure provides, in at leastone embodiment, in accordance with the teachings herein, a chemicalcompound or salt thereof having formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group or anN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group.

In at least one embodiment, in an aspect, R₂ can be an amino group orN-substituted amino group, R₅, R₆ and R₇ can each be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a phosphate group, ahydrogen atom, a hydroxy group, or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₄ can be an amino group orN-substituted amino group and R₂, R₅, R₆ and R₇ can each be a hydrogenatom or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₅ can be an amino group orN-substituted amino group, R₂, R₆ and R₇ can each be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a phosphate group, ahydrogen atom, a hydroxy group, or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₆ can be an amino group orN-substituted amino group, R₂, R₅ and R₇ can each be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a phosphate group, ahydrogen atom, a hydroxy group, or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₇ can be an amino group orN-substituted amino group, R₂, R₅ and R₆ can each be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a phosphate group, ahydrogen atom, a hydroxy group, or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, at least two of R₂, R₄, R₅, R₆or R₇ can be an amino group or N-substituted amino group.

In at least one embodiment, in an aspect, R₂ and R₄ can be an aminogroup or N-substituted amino group, and R₅, R₆ and R₇ can be a hydrogenatom or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₂ and R₅ can be an aminogroup or N-substituted amino group, R₆ and R₇ can be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a hydrogen atom, an alkylgroup, O-alkyl group, a hydroxy group, or a phosphate group.

In at least one embodiment, in an aspect, R₂ and R₆ can be an aminogroup or N-substituted amino group, R₅ and R₇ can be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a hydrogen atom, an alkylgroup, O-alkyl group, a hydroxy group, or a phosphate group.

In at least one embodiment, in an aspect, R₂ and R₇ can be an aminogroup or N-substituted amino group, R₅ and R₆ can be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a hydrogen atom, an alkylgroup an alkyl group, O-alkyl group, a hydroxy group, or a phosphategroup.

In at least one embodiment, in an aspect, R₄ and R₅ can be an aminogroup or N-substituted amino group, and R₂, R₆ and R₇ can be a hydrogenatom or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₄ and R₆ can be an aminogroup or N-substituted amino group, and R₂, R₅ and R₇ can be a hydrogenatom or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₄ and R₇ can be an aminogroup or N-substituted amino group and R₂, R₅ and R₆ can be a hydrogenatom or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₅ and R₆ can be an aminogroup or N-substituted amino group, R₂ and R₇ can be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a hydrogen atom, an alkylgroup, O-alkyl group, a hydroxy group, or a phosphate group.

In at least one embodiment, in an aspect, R₅ and R₇ can be an aminogroup or N-substituted amino group, R₂ and R₆ can be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a hydrogen atom, an alkylgroup, O-alkyl group, a hydroxy group, or a phosphate group.

In at least one embodiment, in an aspect, R₆ and R₇ can be an aminogroup or N-substituted amino group, R₂ and R₅ can be a hydrogen atom oran alkyl group or O-alkyl group, and R₄ can be a hydrogen atom, an alkylgroup, O-alkyl group, a hydroxy group, or a phosphate group.

In at least one embodiment, in an aspect, R₄, R₅, R₆ or R₇ can be anN-substituted amino group.

In at least one embodiment, in an aspect, R₇ can be an N-substitutedamino group.

In at least one embodiment, in an aspect, R₄, R₅, R₆ or R₇ can be anN-substituted amino group, wherein the N-substituted group has theformula:

wherein R′ and R″ are each independently selected from a hydrogen atom,an alkyl group and an aryl group.

In at least one embodiment, in an aspect, R₇ can be an N-substitutedamino group, wherein the N-substituted group has the formula:

wherein R′ and R″ are each independently selected from a hydrogen atom,an alkyl group and an aryl group.

In at least one embodiment, in an aspect, R₄, R₅, R₆ or R₇ can be anN-substituted amino group, wherein the N-substituted group has theformula:

wherein R′ and R″ each are a hydrogen atom.

In at least one embodiment, in an aspect, R₇ can be an N-substitutedamino group, wherein the N-substituted group has the formula:

wherein R′ and R″ each are a hydrogen atom.

In at least one embodiment, in an aspect, R₄ when it is not aminated canbe a hydrogen atom.

In at least one embodiment, in an aspect, R₄ when it is not aminated canbe a hydroxy group.

In at least one embodiment, in an aspect, R₄ when it is not aminated canbe an O-alkyl group.

In at least one embodiment, in an aspect, R₄ when it is not aminated canbe a phosphate group.

In at least one embodiment, in an aspect, three, four or all five of R₂,R₄, R₅, R₆ or R₇ can be an amino group or N-substituted amino group.

In at least one embodiment, in an aspect, the chemical compound can beselected from the group consisting of compounds having formulas (III);(IV); (V); (VI); (VII); (VIII); (IX); (X); (XI); (XII); (XIII); (XIV);(XV); (XVI); and (XVII):

In at least one embodiment, in an aspect, the chemical compound can beat least about 95% (w/w) pure.

In another aspect, the present disclosure relates to pharmaceutical andrecreational drug formulations comprising aminated psilocybinderivatives. Accordingly, in one aspect, the present disclosureprovides, in at least one embodiment, a pharmaceutical or recreationaldrug formulation comprising an effective amount of a chemical compoundor salt thereof having formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group, together with a pharmaceutically acceptable excipient,diluent or carrier.

In another aspect, the present disclosure relates to methods oftreatment of psychiatric disorders. Accordingly, the present disclosurefurther provides, in at least one embodiment, a method for treating apsychiatric disorder, the method comprising administering to a subjectin need thereof a pharmaceutical formulation comprising a chemicalcompound or salt thereof having formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group, wherein the pharmaceutical formulation is administered in aneffective amount to treat the psychiatric disorder in the subject.

In at least one embodiment, in an aspect, the disorder can be a5-HT_(2A) receptor mediated disorder or a 5-HT_(1A) receptor mediateddisorder.

In at least one embodiment, in an aspect, a dose can be administered ofabout 0.001 mg to about 5,000 mg.

In another aspect, the present disclosure relates to methods of makingaminated psilocybin derivatives. Accordingly, in one aspect, the presentdisclosure provides, in at least one embodiment, a method of making anaminated psilocybin derivative the method comprising:

reacting a reactant psilocybin derivative compound or a salt thereofhaving the formula (II):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is a reactive groupselected from a nitro group, an azido group, or a hydrogen atom, andwherein R₂, R₄, R₅, R₆, or R₇ which are not a reactive group, are ahydrogen atom, an alkyl group or O-alkyl group and wherein R_(3A) andR_(3B) are each independently a hydrogen atom, an alkyl group, and acylgroup, or an aryl group under conditions sufficient to form a chemicalcompound having formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group.

In a least one embodiment, in an aspect, R₂, R₄, R₅, R₆, or R₇ in thereactant psilocybin derivative compound can be a hydrogen atom, and theconditions can comprise (i) reacting the reactant psilocybin compoundwith a nitrogenous compound selected from nitric acid (HNO₃); a nitratesalt; an acyl nitrate; trifluoromethansulfonyl nitrate; nitrosoniumtetrafluoroborate (NO₂BF₄); and trifluoracetyl nitrate to form anitrated compound having chemical formula (XXV):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is a nitro group, andwherein each non-nitrated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkylgroup or O-alkyl group, wherein R₄ when it is not nitrated is a hydrogenatom, an alkyl group or O-alkyl group, a hydroxy group, or a phosphategroup, and wherein R_(3A) and R_(3B) are each independently a hydrogenatom, an alkyl group, an aryl group, or an acyl group, and then (ii)reacting the nitrated compound under reducing conditions to form anaminated compound having chemical formula (XXVI):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group, andwherein each non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkylgroup or O-alkyl group, wherein R₄ when it is not aminated is a hydrogenatom, an alkyl group or O-alkyl group, a hydroxy group, or a phosphategroup, and wherein R_(3A) and R_(3B) are each independently a hydrogenatom, an alkyl group, an aryl group, or an acyl group to thereby form acompound having chemical formula (I), wherein at least one of R₂, R₄,R₅, R₆, or R₇ is an amino group, and (iii) optionally substituting theat least one amino group to form at least one N-substituted group.

In at least one embodiment, in an aspect, R₂, R₄, R₅, R₆, or R₇ in thereactant psilocybin derivative compound can be a nitro group or anazide, and the reaction conditions can comprise reacting the reactantpsilocybin compound under reducing conditions to form an aminatedcompound having chemical formula (XXVI):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group, andwherein each non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkylgroup or O-alkyl group, wherein R₄ when it is not aminated is a hydrogenatom, an alkyl group or O-alkyl group, a hydroxy group, or a phosphategroup, and wherein R_(3A) and R_(3B) are each independently a hydrogenatom, an alkyl group, an aryl group, or an acyl group to thereby form acompound having chemical formula (I), wherein at least one of R₂, R₄,R₅, R₆, or R₇ is an amino group, and to then optionally substitute theat least one amino group to form at least one N-substituted group.

In at least one embodiment, in an aspect, at least one of R₂, R₄, R₅,R₆, or R₇ in the reactant psilocybin derivative compound can be ahydrogen atom, and the conditions can comprise reacting the reactantpsilocybin compound with ammonia and hydrogen peroxide in the presenceof a catalyst to form the chemical compound having formula (I) and tothen optionally substitute the at least one amino group in the chemicalcompound having formula (I) to form at least one N-substituted group.

In the least one embodiment, in an aspect, the catalyst can be Cu/SiO₂.

In the least one embodiment, in an aspect, the method can comprise:

-   -   (i) reacting the reactant psilocybin derivative compound to        protect the side-chain amino groups R_(3a) and R_(3b), and,        optionally, protect nitrogen atom N₁ of the reactant psilocybin        derivative compound to obtain an amino-protected compound        wherein R_(3a) and R_(3b) are each a protective group, and        wherein, optionally, N₁ is protected by protective group,    -   (ii) reacting the amino-protected compound with a nitrogenous        compound selected from nitric acid (HNO₃), a nitrate salt, an        acyl nitrate, benzoyl nitrate, nitrosonium tetrafluoroborate        (NO₂BF₄), and trifluoracetyl nitrate to form a nitrated compound        having chemical formula (XXXI):

-   -   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is a nitro            group, and wherein each non-nitrated R₂, R₅, R₆, or R₇ is a            hydrogen atom, an alkyl group or O-alkyl group, wherein R₄            when it is not nitrated is a hydrogen atom, an alkyl group,            O-alkyl group, a hydroxy group, or a phosphate group, and            wherein R_(3a) and R_(3b) are a protective group, and            wherein R₁ is a protective group or a hydrogen atom;

    -   (iii) reacting the compound having chemical formula (XXXI) under        reducing conditions to form a compound having chemical formula        (XXXII):

-   -   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino            group, and wherein each non-aminated R₂, R₅, R₆, or R₇ is a            hydrogen atom, an alkyl group or O-alkyl group, wherein R₄            when it is not aminated is a hydrogen atom, an alkyl group,            O-alkyl group, a hydroxy group, or a phosphate group,            wherein R_(3a) and R_(3b), are a protective group, and            wherein R₁ is a protective group or a hydrogen atom; and

    -   (iv) reacting the compound having chemical formula (XXXII) to        remove the protective groups to thereby form the aminated        compound having chemical formula (I).

In the least one embodiment, in an aspect, the protective group can bean alkyl group, an acyl group, an acetyl group, a substituted acetylgroup, or carbamate group.

In at least one embodiment, in an aspect, the carbamate group can befluorenylmethyloxycarbonyl (Fmoc), benzyloxycarbonyl, ortert-butyloxycarbonyl (Boc).

In the least one embodiment, in an aspect, the method can comprisecomprising performing an additional step following step (iii) and priorto step (iv), the additional step comprising substituting the aminogroup at the at least one R₂, R₄, R₅, R₆, or R₇ groups to form at leastone N-substituted amino group.

In the least one embodiment, in an aspect, the method can compriseperforming an additional step following step (iv), the additional stepcomprising, in a compound wherein R_(3A) and R_(3B) are each a hydrogenatom, substituting at least one of the hydrogen atoms by an alkyl group,an aryl group, or an acyl group form a N-substituted amino group.

In at least one embodiment, in an aspect, R₂ in the compound havingformula (I) can be an amino group or N-substituted amino group, R₅, R₆and R₇ can each be a hydrogen atom or an alkyl group or O-alkyl group,and R₄ can be a phosphate group, a hydrogen atom, a hydroxy group, or analkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₄ in the compound havingformula (I) can be an amino group or N-substituted amino group and R₂,R₅, R₆ and R₇ can each be a hydrogen atom or an alkyl group or O-alkylgroup.

In at least one embodiment, in an aspect, R₅ in the compound havingformula (I) can be an amino group or N-substituted amino group, R₂, R₆and R₇ can each be a hydrogen atom or an alkyl group or O-alkyl group,and R₄ can be a phosphate group, a hydrogen atom, a hydroxy group, or analkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₆ in the compound havingformula (I) can be an amino group or N-substituted amino group atom, R₂,R₅ and R₇ can each be a hydrogen atom or an alkyl group or O-alkylgroup, and R₄ can be a phosphate group, a hydrogen atom, a hydroxygroup, or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, R₇ in the compound havingformula (I) can be an amino group or N-substituted amino group, R₂, R₅and R₆ can each be a hydrogen atom or an alkyl group or O-alkyl group,and R₄ can be a phosphate group, a hydrogen atom, a hydroxy group, or analkyl group or O-alkyl group.

In at least one embodiment, in an aspect, at least two of R₂, R₄, R₅, R₆or R₇ in the compound formula (I) can be an amino group or N-substitutedamino group.

In at least one embodiment, in an aspect, R₂ and R₄ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,and R₅, R₆ and R₇ can be a hydrogen atom or an alkyl group or O-alkylgroup.

In at least one embodiment, in an aspect, R₂ and R₅ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,R₆ and R₇ can be a hydrogen atom, or an alkyl group or O-alkyl group,and R₄ can be a hydrogen atom, an alkyl group, O-alkyl group, a hydroxygroup, or a phosphate group.

In at least one embodiment, in an aspect, R₂ and R₆ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,R₅ and R₇ can be a hydrogen atom or an alkyl group or O-alkyl group, andR₄ can be a hydrogen atom, an alkyl group, O-alkyl group, a hydroxygroup, or a phosphate group.

In at least one embodiment, in an aspect, R₂ and R₇ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,R₅ and R₆ can be a hydrogen atom or an alkyl group or O-alkyl group, andR₄ can be a hydrogen atom, an alkyl group, O-alkyl group, a hydroxygroup, or a phosphate group.

In at least one embodiment, in an aspect, R₄ and R₅ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,and R₂, R₆ and R₇ can be a hydrogen atom, or an alkyl group or O-alkylgroup.

In at least one embodiment, in an aspect, R₄ and R₆ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,and R₂, R₅ and R₇ can be a hydrogen atom or an alkyl group or O-alkylgroup.

In at least one embodiment, in an aspect, R₄ and R₇ in the compoundhaving formula (I) can be an amino group or N-substituted amino groupand R₂, R₅ and R₆ can be a hydrogen atom or an alkyl group or O-alkylgroup.

In at least one embodiment, in an aspect, R₅ and R₆ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,R₂ and R₇ can be a hydrogen atom or an alkyl group or O-alkyl group, andR₄ can be a hydrogen atom, an alkyl group, O-alkyl group, a hydroxygroup, or a phosphate group.

In at least one embodiment, in an aspect, R₅ and R₇ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,R₂ and R₆ can be a hydrogen atom or an alkyl group or O-alkyl group, andR₄ can be a hydrogen atom, an alkyl group, O-alkyl group, a hydroxygroup, or a phosphate group.

In at least one embodiment, in an aspect, R₆ and R₇ in the compoundhaving formula (I) can be an amino group or N-substituted amino group,R₂ and R₅ can be a hydrogen atom or an alkyl group or O-alkyl group, andR₄ can be a hydrogen atom, an alkyl group, O-alkyl group, a hydroxygroup, or a phosphate group.

In at least one embodiment, in an aspect, R₄ in the compound havingformula (I) when it is not aminated can be a hydrogen atom.

In at least one embodiment, in an aspect, R₄ in the compound havingformula (I) when it is not aminated can be a hydroxy group.

In at least one embodiment, in an aspect, R₄ in the compound havingformula (I) when it is not aminated can be an alkyl group.

In at least one embodiment, in an aspect, R₄ when it is not aminated canbe an O-alkyl group.

In at least one embodiment, in an aspect, R₄ in the compound havingformula (I) when it is not aminated can be a phosphate group.

In at least one embodiment, in an aspect, three, four or all five of R₂,R₄, R₅, R₆ or R₇ in the compound having formula (I) can be an aminogroup or N-substituted amino group.

In at least one embodiment, in an aspect, the compound having formula(I) can be selected from the group consisting of compounds havingformulas (Ill); (IV); (V); (VI); (VII); (VIII); (IX); (X); (XI); (XII);(XIII); (XIV); (XV); (XVI); and (XVII):

In another aspect, the present disclosure relates to further methods ofmaking aminated psilocybin derivatives. Accordingly, in one aspect, thepresent disclosure provides in at least one aspect, a method of makingan aminated psilocybin derivative the method comprising:

-   -   (a) contacting an aminated psilocybin precursor compound with a        host cell comprising a psilocybin biosynthetic enzyme        complement; and    -   (b) growing the host cell to produce an aminated psilocybin        derivative or salts thereof having the formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group or anN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group.

In at least one embodiment, in an aspect, the psilocybin biosyntheticenzyme complement can comprise at least one enzyme selected from anucleic acid selected from:

-   -   (a) SEQ. ID NO: 4, SEQ. ID NO: 8, SEQ. ID NO: 11, and SEQ. ID        NO: 13;    -   (b) a nucleic acid sequence that is substantially identical to        any one of the nucleic acid sequences of (a);    -   (c) a nucleic acid sequence that is substantially identical to        any one of the nucleic acid sequences of (a) but for the        degeneration of the genetic code;    -   (d) a nucleic acid sequence that is complementary to any one of        the nucleic acid sequences of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 5, SEQ.        ID NO: 9, SEQ. ID NO: 12, or SEQ. ID NO: 14;    -   (f) a nucleic acid sequence that encodes a functional variant of        any one of the amino acid sequences set forth in SEQ. ID NO: 5,        SEQ. ID NO:7, SEQ. ID NO: 12, or SEQ. ID NO: 14; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f).

In at least one embodiment, in an aspect, the aminated psilocybinprecursor compound can be a compound, having the formula (XXVII):

-   -   wherein at least one of R₂, R₄, R₅, R₆ and R₇ is an amino group        or N-substituted amino group, wherein R₂, R₄, R₅, R₆ and R₇ when        they are not aminated are hydrogen atoms, an alkyl group or        O-alkyl group, wherein R₄ when it is not aminated is a hydrogen        atom, an alkyl group or O-alkyl group, a hydroxy group, or a        phosphate group;        wherein the psilocybin biosynthetic enzyme complement can        comprise:        a tryptophan decarboxylase encoded by a nucleic acid sequence        selected from:    -   (a) SEQ. ID NO: 11;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 12;    -   (f) a nucleic acid sequence that encodes a functional variant of        the amino acid sequence set forth in SEQ. ID NO: 12; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f); and the formed aminated        psilocybin derivative can be a compound having formula (XXVIII):

-   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group        or an N-substituted amino group, and wherein each non-aminated        R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkyl group, or O-alkyl        group, wherein R₄ when it is not aminated is a hydrogen atom, an        alkyl or O-alkyl group, a hydroxy group, or a phosphate group,        and wherein at least one of R_(3A) and R_(3B) are each        independently a hydrogen atom.

In at least one embodiment, in an aspect, the aminated psilocybinprecursor compound can be an aminated indole compound having the formula(XXIX):

-   -   wherein at least one of R₂, R₄, R₅, R₆ and R₇ is an amino group        or N-substituted amino group, wherein R₂, R₄, R₅, R₆ and R₇ when        they are not aminated are hydrogen atoms, an alkyl group or        O-alkyl group, wherein R₄ when it is not aminated is a hydrogen        atom, an alkyl group or O-alkyl group, a hydroxy group, or a        phosphate group;        wherein the psilocybin biosynthetic enzyme complement can        comprise:

(i) a tryptophan synthase subunit B polypeptide encoded by a nucleicacid selected from:

-   -   (a) SEQ. ID NO: 8;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 9;    -   (f) a nucleic acid sequence that encodes a functional variant of        the amino acid sequence set forth in SEQ. ID NO: 9; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f); and

(ii) a tryptophan decarboxylase encoded by a nucleic acid sequenceselected from:

-   -   (a) SEQ. ID NO: 11;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 12;    -   (f) a nucleic acid sequence that encodes a functional variant of        the amino acid sequence set forth in SEQ. ID NO: 12; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to

any one of the nucleic acid sequences set forth in (a), (b), (c), (d),(e) or (f); and wherein the formed aminated psilocybin derivative can bea compound having formula (XXVIII):

-   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group        or an N-substituted amino group, and wherein each non-aminated        R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkyl group or O-alkyl        group, wherein R₄ when it is not aminated is a hydrogen atom, an        alkyl group or O-alkyl group, a hydroxy group, or a phosphate        group, and wherein at least one of R_(3A) and R_(3B) are        hydrogen atom.

In at least one embodiment, in an aspect, R_(3A) and R_(3B) in formula(XXVIII) can each be a hydrogen atom.

In at least one embodiment, in an aspect, the psilocybin biosyntheticenzyme complement can further comprise an N-acetyl transferase.

In at least one embodiment, in an aspect, the N-acetyl transferase canbe an enzyme encoded by. a nucleic acid sequence selected from:

-   -   (a) SEQ. ID NO: 4;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 5;    -   (f) a nucleic acid sequence that encodes a functional variant of        the amino acid sequence set forth in SEQ. ID NO: 5; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f).

In at least one embodiment, in an aspect, the formed aminated psilocybincompound can have the formula (XXX):

-   -   wherein at least one of R₂, R₄, R₅, R₆ or R₇ is an amino group        or substituted amino group, wherein each non-aminated R₂, R₅,        R₆, or R₇ is a hydrogen atom, or an alkyl group or O-alkyl        group, wherein R₄ when it is not aminated is a phosphate group,        a hydrogen atom or an alkyl group or O-alkyl group.

In at least one embodiment, in an aspect, the psilocybin biosyntheticenzyme complement can further comprise an N-methyl transferase.

In at least one embodiment, in an aspect, the N-methyl transferase canbe an enzyme encoded by a nucleic acid sequence selected from:

-   -   (a) SEQ. ID NO: 13;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 14;    -   (f) a nucleic acid sequence that encodes a functional variant of        the amino acid sequence set forth in SEQ. ID NO: 14; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f).

In at least one embodiment, in an aspect, the formed aminated psilocybincompound can have the chemical formula (XXXIII):

-   -   wherein at least one of R₂, R₄, R₅, R₆ or R₇ is an amino group        or substituted amino group, wherein each non-aminated R₂, R₅,        R₆, or R₇ is a hydrogen atom, or an alkyl group or O-alkyl        group, wherein R₄ when it is not aminated is a phosphate group,        a hydrogen atom or an alkyl group or O-alkyl group, and wherein        at least one of R_(3a) and R_(3b) is an amino group, and wherein        a non-aminated R_(3a) and R_(3b) is a hydrogen atom.

In at least one embodiment, in an aspect, the aminated psilocybinderivative compound having formula (I) can be selected from the groupconsisting of compounds having formulas (Ill); (IV); (V); (VI); (VII);(VIII); (IX); (X); (XI); (XII); (XIII); (XIV); (XV); (XVI); and (XVII):

In at least one embodiment, in an aspect, the aminated psilocybinprecursor compound can be contacted with the host cell by including theaminated psilocybin precursor compound in a growth medium for the hostcell.

In at least one embodiment, in an aspect, the method can further includea step comprising isolating the aminated psilocybin derivative.

In at least one embodiment, in an aspect, the host cell can be amicrobial cell.

In at least one embodiment, in an aspect, the host cell can be abacterial cell or a yeast cell.

In another aspect, the present disclosure provides, in at least oneembodiment, a method for modulating a 5-HT_(2A) receptor or a 5-HT_(1A)receptor, the method comprising contacting the 5-HT_(2A) receptor or the5-HT_(1A) receptor with a chemical compound or salts thereof having theformula (I):

under conditions sufficient to modulate the 5-HT_(2A) receptor or the5-HT_(1A) receptor, wherein at least one of R₂, R₄, R₅, R₆, or R₇ is anamino group or N-substituted amino group, and wherein each non-aminatedR₂, R₅, R₆, or R₇ is a hydrogen atom, an alkyl group or O-alkyl group,wherein R₄ when it is not aminated is a hydrogen atom, an alkyl group orO-alkyl group, a hydroxy group, or a phosphate group, and wherein R_(3A)and R_(3B) are each independently a hydrogen atom, an alkyl group, anaryl group, or an acyl group, together with a pharmaceuticallyacceptable excipient, diluent or carrier.

In at least one embodiment, in an aspect, the reaction conditions can bein vitro reaction conditions.

In at least one embodiment, in an aspect, the reaction conditions can bein vivo reaction conditions.

In another aspect, the present disclosure provides, in at least oneembodiment, a use of a chemical compound having the formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a phosphate group, a hydrogen atom, or a hydroxygroup, an alkyl group or O-alkyl group and wherein R_(3A) and R_(3B) area hydrogen atom, an alkyl group or O-alkyl group, in the manufacture ofa pharmaceutical or recreational drug formulation.

In at least one embodiment, in an aspect, the manufacture can compriseformulating the chemical compound with an excipient, diluent or carrier.

In at least one embodiment, in an aspect, the manufacture can furtherinclude a step comprising derivatizing the chemical compound having theformula (I) by substituting the amino group or N-substituted amino groupwith another group or an atom.

In another aspect, the present disclosure provides, in at least oneembodiment, a use of a chemical compound having the formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a phosphate group, a hydrogen atom, a hydroxygroup, or an alkyl group or O-alkyl group, and wherein R_(3A) and R_(3B)are each independently a hydrogen atom, an alkyl group or O-alkyl group,together with a diluent, carrier, or excipient as a pharmaceutical orrecreational drug formulation.

Other features and advantages will become apparent from the followingdetailed description. It should be understood, however, that thedetailed description, while indicating preferred implementations of thedisclosure, are given by way of illustration only, since various changesand modifications within the spirit and scope of the disclosure willbecome apparent to those of skill in the art from the detaileddescription.

BRIEF DESCRIPTION OF THE DRAWINGS

The disclosure is in the hereinafter provided paragraphs described, byway of example, in relation to the attached figures. The figuresprovided herein are provided for a better understanding of the exampleembodiments and to show more clearly how the various embodiments may becarried into effect. The figures are not intended to limit the presentdisclosure.

FIG. 1 depicts the chemical structure of psilocybin.

FIG. 2 depicts a certain prototype structure of psilocybin andpsilocybin derivative compounds, namely an indole. Certain carbon andnitrogen atoms may be referred to herein by reference to their positionwithin the indole structure, i.e., N₁, C₂, C₃ etc. The pertinent atomnumbering is shown.

FIGS. 3A, 3B, 3C, 3D, 3E, 3F, 3G, 3H, 3I, 3J, 3K, 3L, 3M, 3N, 3O, and 3Pdepict the chemical structures of certain example aminated psilocybinderivatives, notably a 2-amino-psilocybin derivative (FIG. 3A), a5-amino-psilocybin derivative (FIG. 3B), a 6-amino-psilocybin derivative(FIG. 3C), a 7-amino-psilocybin derivative (FIG. 3D), a2-N,N-substituted-amino-psilocybin derivative (FIG. 3E), a5-N,N-substituted-amino-psilocybin derivative (FIG. 3F), a6-N,N-substituted-amino-psilocybin derivative (FIG. 3G), a7-N,N-substituted amino-psilocybin derivative (FIG. 3H), a2-amino-6-ethyl-psilocybin derivative (FIG. 3I), a 5-amino-7-methylpsilocybin derivative (FIG. 3J), a 5-O-ethyl-6-amino-psilocybinderivative (FIG. 3K), a 6-O-methyl-7-amino-psilocybin (FIG. 3L), a2-N,N-substituted amino-7-methyl-psilocybin derivative (FIG. 3M), a5-N,N-substituted amino-6-ethyl-psilocybin derivative (FIG. 3N), a5-O-methyl-6-N,N-substituted amino-psilocybin derivative (FIG. 3O), anda 5-O-ethyl-7-N,N-substituted amino-psilocybin derivative (FIG. 3P).

FIGS. 4A, 4B, 4C, 4D, 4E, 4F, 4G, 4H, 4I, 4J, 4K, 4L, 4M, 4N, 4O, 4P,4Q, 4R, 4S, 4T, 4U, and 4V depict the chemical structures of certainexample aminated psilocybin derivatives, notably alkylatedhydroxy-containing psilocybin derivatives, notably a2,5-di-amino-psilocybin derivative (FIG. 4A), a 2,6-di-amino-psilocybinderivative (FIG. 4B), a 2,7-di-amino-psilocybin derivative (FIG. 4C), a5,6-di-amino-psilocybin derivative (FIG. 4D), a 5,7-di-amino-psilocybinderivative (FIG. 4E), a 6,7-di-amino-psilocybin derivative (FIG. 4F), a2,5,6-tri-amino-psilocybin derivative (FIG. 4G), a2,5,7-tri-amino-psilocybin derivative (FIG. 4H), a2,6,7-tri-amino-psilocybin derivative (FIG. 4I) a5,6,7-tri-amino-psilocybin derivative (FIG. 4J), a2,5,6,7-tetra-amino-psilocybin derivative (FIG. 4K), a2-N,N,5-N,N-di-substituted-amino-psilocybin derivative (FIG. 4L), a2-N,N,6-N,N-di-substituted-amino-psilocybin derivative (FIG. 4M), a2-N,N,7-N,N-di-substituted-amino-psilocybin derivative (FIG. 4N), a5-N,N,6-N,N-di-substituted-amino-psilocybin derivative (FIG. 4O), a5-N,N,7-N,N-disubstituted-amino-psilocybin derivative (FIG. 4P), a6-N,N,7-N,N-di-substituted-amino-psilocybin derivative (FIG. 4Q), a2-N,5-N,6-N,N-tri-substituted-amino-psilocybin derivative (FIG. 4R), a2-N,N,5-N,N,7-N,N-tri-substituted-amino-psilocybin derivative (FIG. 4S),a 2-N,N,6-N,N,7-N,N-tri-substituted-amino-psilocybin derivative (FIG.4T) a 5-N,N,6-N,N,7-N,N-tri-substituted-amino-psilocybin derivative(FIG. 4U), a 2-N,N,5-N,N,6-N,N,7-N,N-tetra-substituted-amino-psilocybinderivative (FIG. 4V).

FIGS. 5A, 5B, 5C, 5D, 5E, 5F, and 5G depict the chemical structures ofcertain example aminated psilocybin derivatives, notably O-alkylatedaminated psilocybin derivatives, notably a 4-O-methyl-5-amino-psilocybinderivative (FIG. 5A), a 4-O-ethyl-5-amino-psilocybin derivative (FIG.5B), O-acylated aminated psilocybin derivatives, notably a4-acetyl-5-amino-psilocybin derivative (FIG. 5C), a4-propanoyl-5-amino-psilocybin derivative (FIG. 5D), a4-hydroxy-5-amino-psilocybin derivative (FIG. 5E), and a4-phospho-5-amino-psilocybin derivative (FIG. 5F), and 4- and a5-amino-psilocybin derivative (FIG. 5G).

FIGS. 6A, 6B, 6C, 6D, 6E, 6F, and 6G depict the chemical structures ofcertain example aminated psilocybin derivatives, notably O-alkylatedaminated psilocybin derivatives, notably a 4-O-methyl-7-amino-psilocybinderivative (FIG. 6A), a 4-O-ethyl-7-amino-psilocybin derivative (FIG.6B), O-acylated aminated psilocybin derivatives, notably a4-acetyl-7-amino-psilocybin derivative (FIG. 6C), a4-propanoyl-7-amino-psilocybin derivative (FIG. 6D), a4-hydroxy-7-amino-psilocybin derivative (FIG. 6E), a4-phospho-7-amino-psilocybin derivative (FIG. 6F), and a7-amino-psilocybin derivative (FIG. 6G).

FIGS. 7A, 7B, 7C, 7D, 7E, 7F, and 7G depict the chemical structures ofcertain example aminated psilocybin derivatives, notably O-alkylatedaminated psilocybin derivatives, notably a4-O-methyl-5,7-di-amino-psilocybin derivative (FIG. 7A), a4-O-ethyl-5,7-di-amino-psilocybin derivative (FIG. 78 ), O-acylatedaminated psilocybin derivatives, notably a4-acetyl-5,7-di-amino-psilocybin derivative (FIG. 7C), a4-propanoyl-5,7-di-amino-psilocybin derivative (FIG. 7D), a4-hydroxy-5,7-di-amino-psilocybin derivative (FIG. 7E), a4-phospho-5,7-di-amino-psilocybin derivative (FIG. 7F), and a5,7-di-amino-psilocybin derivative (FIG. 7G).

FIGS. 8A, 8B, 8C, 8D, 8E, 8F, and 8G depict the chemical structures ofcertain example psilocybin derivatives, notably O-alkylated psilocybinderivatives, notably a 4-O-methyl-psilocybin derivative (FIG. 8A), a4-O-ethyl-psilocybin derivative (FIG. 8B), a 4-acetyl-psilocybinderivative (FIG. 8C), a 4-propanoyl-psilocybin derivative (FIG. 8D), a4-hydroxy-psilocybin derivative (FIG. 8E), a 4-phospho-psilocybinderivative (FIG. 8F), and a 4-psilocybin derivative (FIG. 8G).

FIGS. 9A, 9B, 9C and 9D depict certain example chemical reactions forsynthesizing aminated psilocybin derivatives with subsequentN-substitutions, notably a reaction wherein a4-O-methyl-5-nitro-psilocybin derivative is reacted with hydrogen underthe catalysis of palladium on charcoal to form a4-O-methyl-5-amino-psilocybin derivative (FIG. 9A). The formed aminogroup at the 5-position can then be substituted with different groupsuch as an acylation with acetic anhydride. The amino group can also bealkylated via a condensation with an aldehyde (such as acetaldehyde)followed by a reduction of the intermediate imine with borohydride (FIG.9B). FIG. 9C depicts a possible direction amination method with H₂O₂ andNH₃.H₂O with the help of a catalyst. FIG. 9D further depicts a multistepsynthesis of two 4-O-methyl-psilocybin derivatives respectively aminatedat C₅ (compound 9D-8) and C₇ (compound 9D-10).

FIG. 10 depicts example biosynthesis processes for the synthesis ofseveral example aminated psilocybin derivatives.

FIGS. 11A, 11B, 11C, 11D, 11E, 11F, 11G, 11H, 11I, 11J, 11K, 11L, and11M depict various graphs, obtained in the performance of experimentalassays to evaluate the efficacy of an example aminated psilocybinderivative having the chemical formulae (IX), (XII) and (XIII) set forthherein, notably a cell viability assay for an aminated psilocybinderivative having the chemical formulae (IX) (FIG. 11A), (XII) (FIG.11B) and (XIII) (FIG. 11C); a saturation binding assay for[³H]ketanserin at the 5-HT_(2A) receptor (FIG. 11D); a competition assayfor psilocin as a positive control (binding) (top panel), and acompetition assay for tryptophan as a negative control (no binding)(bottom panel) (FIG. 11E); a competition assay for an aminatedpsilocybin derivative compound with formula (IX), designated “IX” (FIG.11F); a competition assay for an aminated psilocybin derivative compoundwith formula (XII), designated “XII” (FIG. 11G); a competition assay foran aminated psilocybin derivative compound with formula (XIII),designated “XIII” (FIG. 11H); a luminescence assay in +5HT_(1A) cellcultures at various forskolin concentrations (FIG. 11I), a luminescenceassay in +5HT_(1A) cell cultures in the presence of constant (4 μM)forskolin but with decreasing DMT concentration (FIG. 11J), aluminescence assay in +5HT_(1A) cell cultures in the presence ofconstant (4 μM) forskolin but with decreasing tryptophan concentration(FIG. 14K), a cAMP assay in the presence of constant (4 μM) forskolinbut with decreasing serotonin concentration in +5HT_(1A) cells (FIG.11L), a cAMP assay in the presence of constant (4 μM) forskolin but withincreasing concentration of an aldehyde psilocybin compound havingformula (IX), designated “IX” in +5HT_(1A) cells and −5HT_(1A) cells(FIG. 11M).

FIGS. 12A and 12B depict a representation of mass spectrometry data inthe form of a chromatogram, notably a chromatogram obtained in theperformance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (VI) setforth herein (FIG. 12A); and a representation of mass spectrometry datain the form of a mass spectrometry spectrum obtained in the performanceof an experiment to identify an aminated psilocybin derivative compoundhaving the chemical formula (VI) set forth herein (FIG. 12B).

FIGS. 13A and 13B depict a representation of further mass spectrometrydata in the form of a chromatogram, notably a chromatogram obtained inthe performance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (XIV) setforth herein (FIG. 13A); and a representation of mass spectrometry datain the form of a further mass spectrometry spectrum obtained in theperformance of an experiment to identify an aminated psilocybinderivative compound having the chemical formula (XIV) set forth herein(FIG. 13B).

FIGS. 14A and 14B depict a representation of further mass spectrometrydata in the form of a chromatogram, notably a chromatogram obtained inthe performance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (VII) setforth herein (FIG. 14A); and a representation of mass spectrometry datain the form of a further mass spectrometry spectrum obtained in theperformance of an experiment to identify an aminated psilocybinderivative compound having the chemical formula (VII) set forth herein(FIG. 14B).

FIGS. 15A and 15B depict a representation of further mass spectrometrydata in the form of a chromatogram, notably a chromatogram obtained inthe performance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (IV) setforth herein (FIG. 15A); and a representation of mass spectrometry datain the form of a further mass spectrometry spectrum obtained in theperformance of an experiment to identify an aminated psilocybinderivative compound having the chemical formula (IV) set forth herein(FIG. 15B).

FIGS. 16A and 16B depict a representation of further mass spectrometrydata in the form of a chromatogram, notably a chromatogram obtained inthe performance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (III) setforth herein (FIG. 16A); and a representation of mass spectrometry datain the form of a further mass spectrometry spectrum obtained in theperformance of an experiment to identify an aminated psilocybinderivative compound having the chemical formula (III) set forth herein(FIG. 16B).

FIGS. 17A and 17B depict a representation of further mass spectrometrydata in the form of a chromatogram, notably a chromatogram obtained inthe performance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (XV) setforth herein (FIG. 17A); and a representation of mass spectrometry datain the form of a further mass spectrometry spectrum obtained in theperformance of an experiment to identify an aminated psilocybinderivative compound having the chemical formula (XV) set forth herein(FIG. 17B).

FIGS. 18A and 18B depict a representation of further mass spectrometrydata in the form of a chromatogram, notably a chromatogram obtained inthe performance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (XVI) setforth herein (FIG. 18A); and a representation of mass spectrometry datain the form of a further mass spectrometry spectrum obtained in theperformance of an experiment to identify an aminated psilocybinderivative compound having the chemical formula (XVI) set forth herein(FIG. 18B).

FIG. 19 depicts a representation of further mass spectrometry data inthe form of a chromatogram, notably a chromatogram obtained in theperformance of an experiment to synthesize an example aminatedpsilocybin derivative compound having the chemical formula (XVII) setforth herein (FIG. 19 ).

The figures together with the following detailed description makeapparent to those skilled in the art how the disclosure may beimplemented in practice.

DETAILED DESCRIPTION

Various compositions, systems or processes will be described below toprovide an example of an embodiment of each claimed subject matter. Noembodiment described below limits any claimed subject matter and anyclaimed subject matter may cover processes, compositions or systems thatdiffer from those described below. The claimed subject matter is notlimited to compositions, processes or systems having all of the featuresof any one composition, system or process described below or to featurescommon to multiple or all of the compositions, systems or processesdescribed below. It is possible that a composition, system, or processdescribed below is not an embodiment of any claimed subject matter. Anysubject matter disclosed in a composition, system or process describedbelow that is not claimed in this document may be the subject matter ofanother protective instrument, for example, a continuing patentapplication, and the applicant(s), inventor(s) or owner(s) do not intendto abandon, disclaim or dedicate to the public any such subject matterby its disclosure in this document.

As used herein and in the claims, the singular forms, such “a”, “an” and“the” include the plural reference and vice versa unless the contextclearly indicates otherwise. Throughout this specification, unlessotherwise indicated, “comprise,” “comprises” and “comprising” are usedinclusively rather than exclusively, so that a stated integer or groupof integers may include one or more other non-stated integers or groupsof integers.

Various compositions, systems or processes will be described below toprovide an example of an embodiment of each claimed subject matter. Noembodiment described below limits any claimed subject matter and anyclaimed subject matter may cover processes, compositions or systems thatdiffer from those described below. The claimed subject matter is notlimited to compositions, processes or systems having all of the featuresof any one composition, system or process described below or to featurescommon to multiple or all of the compositions, systems or processesdescribed below. It is possible that a composition, system, or processdescribed below is not an embodiment of any claimed subject matter. Anysubject matter disclosed in a composition, system or process describedbelow that is not claimed in this document may be the subject matter ofanother protective instrument, for example, a continuing patentapplication, and the applicant(s), inventor(s) or owner(s) do not intendto abandon, disclaim or dedicate to the public any such subject matterby its disclosure in this document.

When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and sub-combinations of ranges and specific embodimentstherein are intended to be included. Other than in the operatingexamples, or where otherwise indicated, all numbers expressingquantities of ingredients or reaction conditions used herein should beunderstood as modified in all instances by the term “about.” The term“about” when referring to a number or a numerical range means that thenumber or numerical range referred to is an approximation withinexperimental variability (or within statistical experimental error), andthus the number or numerical range may vary between 1% and 15% of thestated number or numerical range, as will be readily recognized bycontext. Furthermore any range of values described herein is intended tospecifically include the limiting values of the range, and anyintermediate value or sub-range within the given range, and all suchintermediate values and sub-ranges are individually and specificallydisclosed (e.g., a range of 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.90, 4,and 5). Similarly, other terms of degree such as “substantially” and“approximately” as used herein mean a reasonable amount of deviation ofthe modified term such that the end result is not significantly changed.These terms of degree should be construed as including a deviation ofthe modified term if this deviation would not negate the meaning of theterm it modifies.

Unless otherwise defined, scientific and technical terms used inconnection with the formulations described herein shall have themeanings that are commonly understood by those of ordinary skill in theart. The terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to limit the scope ofthe present invention, which is defined solely by the claims.

All publications, patents and patent applications are hereinincorporated by reference in their entirety to the same extent as ifeach individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by referencein its entirety.

Terms and Definitions

The term “psilocybin”, refers to a chemical compound having thestructure set forth in FIG. 1 .

The term “indole prototype structure” refers to the chemical structureshown in FIG. 2 . It is noted that specific carbon atoms and a nitrogenatom in the indole prototype structure are numbered. Reference may bemade to these carbon and nitrogen numbers herein, for example C₂, C₄,N₁, and so forth. Furthermore, reference may be made to chemical groupsattached to the indole prototype structure in accordance with the samenumbering, for example R₄ and R₆ reference chemical groups attached tothe C₄ and C₆ atom, respectively. In addition, R_(3A) and R_(3B), inthis respect, reference chemical groups extending from the ethyl-aminogroup extending in turn from the C₃ atom of the prototype indolestructure.

The term “aminated psilocybin derivative” refers to a psilocybinderivative compound to which an amino group has been bonded topsilocybin or a psilocybin derivative. The nitrogen of the amino groupmay bear 1-3 substituents (i.e., be a N-substituted amino group).N-substituents can be an alkyl, aryl, acyl, sulfonyl groups orcombinations thereof. Reference may be made to specific carbon atomswhich may be aminated. For example, a 5-amino-psilocybin derivativerefers to a psilocybin derivative in which carbon atom number 5 (asidentified in the indole prototype structure) possesses an amino orN-substituted amino group, or, similarly, 7-amino-psilocybin derivativerefers to a psilocybin derivative in which carbon atom number 7 (asidentified in the indole prototype structure) possess an amino orN-substituted amino group. Thus, for example, aminated psilocybinderivatives include, single amino derivatives, 2-amino, 4-amino,5-amino, 6-amino, and 7-amino psilocybin derivatives, for example, andmultiple amino derivatives, such as, for example, 5,7-di-aminopsilocybin derivatives, and 2,5,7-tri-amino psilocybin derivatives. Theterm aminated psilocybin derivatives further includes chemical compoundshaving the chemical formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group or anN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom or an alkyl group or O-alkyl group, wherein R₄when it is not aminated is a hydrogen atom, an alkyl group or O-alkylgroup, a hydroxy group, or a phosphate group, and wherein R_(3A) andR_(3B) are a hydrogen atom, an alkyl group, an aryl group, or an acylgroup. Furthermore, it is noted that when R₄ is a phosphate group, theterm aminated psilocybin derivatives includes compounds having theformula (XIX):

wherein at least one of R₂, R₅, R₆, or R₇ is a hydrogen atom, andwherein R₂, R₅, R₆, or R₇ which are not a hydrogen atom are an amino orN-substituted amino group, and wherein R_(3A) and R_(3B) are a hydrogenatom, an alkyl group or aryl group. The term further includes salts ofaminated psilocybins, such as a sodium salt, a potassium salt etc.

The term “amino group” and “amino”, as used herein refers to a moleculecontaining one atom of nitrogen bonded to hydrogen atoms and having theformula —NH₂. An amino group also may be protonated and having theformula —NH₃ ⁺. An amino group through its nitrogen atom may bechemically bonded to another entity. Furthermore, it is noted that anentity attached to an amino group may be referred to herein as an“aminated” entity, e.g., an aminated psilocybin derivative is apsilocybin derivative possessing either an amino group or aN-substituted amino group.

The term “N-substituted amino group”, as used herein, refers to an aminogroup wherein at least one of the hydrogen atoms has been substituted byanother atom or group, such as, for example, an alkyl group, an acylgroup, an aryl group a sulfonyl group etc., excluding, however, an aminogroup wherein both of the hydrogen atoms are substituted by oxygen atomsto thereby form a nitro group. An N-substituted amino group also may beprotonated, and the amino group through its nitrogen atom may bechemically bonded to another entity. Thus, N-substituted amino group maybe represented herein as:

Furthermore N-substituted amino groups include:

-   -   (i) chemical group (XX) (an alkyl group, an aryl group):

wherein R′ and R″ are each independently selected from a hydrogen atom,an alkyl group, and an aryl group, provided however that at least one ofR′, and R″ is not a hydrogen atom;

-   -   (ii) chemical group (XXI):

wherein R′, R″ and R′″ are each independently selected from a hydrogenatom, an alkyl group, and an aryl group, provided however that at leastone of R′, R″, and R′″ is not a hydrogen atom;

-   -   (iii) chemical group (XXII) (an acyl group):

wherein R′ and R″ are each independently selected from a hydrogen atom,an alkyl group and an aryl group;

-   -   (iv) chemical group (XXIII) (a sulfonyl group):

wherein R′, and R″ are each independently selected from a hydrogen atom,an alkyl group and an aryl group; or

-   -   (v) chemical group (XXIV) (a sulfonate group):

wherein R″ is selected from a hydrogen atom, an alkyl group, and an arylgroup. The nitrogen atom of chemical groups (XXII), (XXIII) and (XXIV)can also be positively charged and be further substituted with H, orR′″.

It is noted that R′, R″ and R′″ can herein additionally includenumerical subscripts, such as 5a, 6b, 7b etc., and be represented, forexample, as R′_(5a), R″_(6b) or R′″_(7a), respectively. Where suchnumerical values are included, they reference chemical entity extendingfrom the amino group extending in turn from the thus numbered C atom ofthe prototype indole structure. Thus, for example, R′_(5a) is a chemicalentity extending from an aminated group attached to the C₅ atom of theindole ring structure, R′_(2a) is a chemical entity extending from anaminated group attached to the C₂ atom of the indole ring structure, andso forth. Furthermore, it is noted that an entity attached to anN-substituted amino group may be referred to herein as an “aminated”entity, e.g., an aminated psilocybin derivative is a psilocybinderivative possessing either an amino group or a N-substituted aminogroup.

The term “sulfonyl”, as used herein, refers to a molecule containing onesulfur atom bonded to two oxygen atoms, and one other entity and havingthe formula:

wherein R may be a variety of entities including a hydroxy group, analkyl group, or an aryl group. A sulfonyl group through its sulfur atommay be chemically bonded to another entity. Furthermore, it is notedthat an entity attached to a sulfonyl group may be referred to herein asa “sulfonylated”.

The term “phosphate group”, as used herein, is a molecule containing oneatom of phosphorus, covalently bound to four oxygen atoms (three singlebonds and one double bond). Of the four oxygen atoms, one oxygen atommay be a hydroxy, and one of the non-hydroxylated oxygen atom may bechemically bonded to another entity.

The terms “hydroxy group”, and “hydroxy”, as used herein, refer to amolecule containing one atom of oxygen bonded to one atom of hydrogen,and having the formula —OH. A hydroxy through its oxygen atom may bechemically bonded to another entity.

The terms “nitro” and “nitro group”, as used herein, refer to a moleculecontaining one atom of nitrogen bonded to two atoms of oxygen and havingthe formula —NO₂. A nitro group through its nitrogen atom may bechemically bonded to another entity. Furthermore, it is noted that anentity attached to a nitro group may be referred to herein as a“nitrated” entity, e.g., a nitrated psilocybin derivative is apsilocybin derivative possessing a nitro group.

The term “alkyl”, as used herein, refers to a straight and/or branchedchain, saturated alkyl radical containing from one to “p” carbon atoms(“C₁-C_(p)-alkyl”) and includes, depending on the identity of “p”,methyl, ethyl, propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl,2,2-dimethylbutyl, n-pentyl, 2-methylpentyl, 3-methylpentyl,4-methylpentyl, n-hexyl and the like, where the variable p is an integerrepresenting the largest number of carbon atoms in the alkyl radicalAlkyl groups further include hydrocarbon groups arranged in a chainhaving the chemical formula —C_(n)H_(2n+1), including, withoutlimitation, methyl groups (—CH₃), ethyl groups (—C₂H₅), propyl groups(—C₃H₇), and butyl groups (—C₄H₉).

The term “O-alkyl”, as used herein, refers to a hydrocarbon grouparranged in a chain having the chemical formula —O—C_(n)H_(2n+1).O-alkyl groups include, without limitation, O-methyl groups (—O—CH₃),O-ethyl groups (—O—C₂H₅), O-propyl groups (—O—C₃H₇) and O-butyl groups(—O—C₄H₉).

The term “aryl”, as used herein, refers to a monocyclic, bicyclic, ortricyclic aromatic ring system containing, depending on the number ofatoms in the rings, for example, from 6 to 14 carbon atoms (C₆-C₁₄-aryl)or from 6 to 10 carbons (C₆-C₁₀-aryl), and at least 1 aromatic ring andincludes phenyl, naphthyl, anthracenyl, 1,2-dihydronaphthyl,1,2,3,4-tetrahydronaphthyl, fluorenyl, phenanthrenyl, biphenylenyl,indanyl, indenyl and the like.

The term “acyl”, as used herein, refers to a carbon atom double bondedto an oxygen and single bonded to an alkyl group. The carbon atomfurther can be bonded to another entity. An acyl group can be describedby the chemical formula: —C(═O)—C_(n)H_(2n+1).

The term “O-acyl group” refers to an acyl group in which the carbon atomis single bonded to an additional oxygen atom. The additional oxygenatom can be bonded to another entity. An O-acyl group can be describedby the chemical formula: —O—C(═O)—C_(n)H_(2n+1). Furthermore, dependingon the carbon chain, length specific O-acyl groups may be termed anacetyl group (n=1), a propanoyl group (n=2), propoxycarbonyl group(n=3), a butoxycarbonyl group (n=4) etc.

The term “azido”, as used herein refers to a chemical group having theformula: —N═N⁺═N⁻.

The term “5-HT_(2A) receptor”, as used herein, refers to a subclass of afamily of receptors for the neurotransmitter and peripheral signalmediator serotonin. 5-HT_(2A) receptors can mediate a plurality ofcentral and peripheral physiologic functions of serotonin. Centralnervous system effects can include mediation of hallucinogenic effectsof hallucinogenic compounds.

The term “modulating 5-HT_(2A) receptors”, as used herein, refers to theability of a compound disclosed herein to alter the function of5-HT_(2A) receptors. A 5-HT_(2A) receptor modulator may activate theactivity of a 5-HT_(2A) receptor, may activate or inhibit the activityof a 5-HT_(2A) receptor depending on the concentration of the compoundexposed to the 5-HT_(2A) receptor, or may inhibit the activity of a5-HT_(2A) receptor. Such activation or inhibition may be contingent onthe occurrence of a specific event, such as activation of a signaltransduction pathway, and/or maybe manifest only in particular celltypes. The term “modulating 5-HT_(2A) receptors,” also refers toaltering the function of a 5-HT_(2A) receptor by increasing ordecreasing the probability that a complex forms between a 5-HT_(2A)receptor and a natural binding partner to form a multimer. A 5-HT_(2A)receptor modulator may increase the probability that such a complexforms between the 5-HT_(2A) receptor and the natural binding partner,may increase or decrease the probability that a complex forms betweenthe 5-HT_(2A) receptor and the natural binding partner depending on theconcentration of the compound exposed to the 5-HT_(2A) receptor, and ormay decrease the probability that a complex forms between the 5-HT_(2A)receptor and the natural binding partner. Furthermore, the term includesallosteric modulation of the receptor 5-HT_(2A), i.e., modulation of the5-HT_(2A) receptor through interaction with the 5-HT_(2A) receptor thatis topographically different than the orthosteric site recognized by thecell's endogenous agonist, such modulation further including positiveallosteric modulation (PAM), negative allosteric modulation (NAM) andsilent allosteric modulation (SAM).

The term “5-HT_(2A) receptor-mediated disorder”, as used herein, refersto a disorder that is characterized by abnormal 5-HT_(2A) receptoractivity. A 5-HT_(2A) receptor-mediated disorder may be completely orpartially mediated by modulating 5-HT_(2A) receptors. In particular, a5-HT_(2A) receptor-mediated disorder is one in which modulation of5-HT_(2A) receptors results in some effect on the underlying disordere.g., administration of a 5-HT_(2A) receptor modulator results in someimprovement in at least some of the subjects being treated.

The term “5-HT_(1A) receptor”, as used herein, refers to a subclass of afamily of receptors for the neurotransmitter and peripheral signalmediator serotonin. 5-HT_(1A) receptors can mediate a plurality ofcentral and peripheral physiologic functions of serotonin. Ligandactivity at 5-HT_(1A) is generally not associated with hallucination,although many hallucinogenic compounds are known to modulate 5-HT_(1A)receptors to impart complex physiological responses (Inserra et al.,2020, Pharmacol Rev 73: 202).

The term “modulating 5-HT_(1A) receptors”, as used herein, refers to theability of a compound disclosed herein to alter the function of5-HT_(1A) receptors. A 5-HT_(2A) receptor modulator may activate theactivity of a 5-HT_(1A) receptor, may activate or inhibit the activityof a 5-HT_(1A) receptor depending on the concentration of the compoundexposed to the 5-HT_(1A) receptor, or may inhibit the activity of a5-HT_(1A) receptor. Such activation or inhibition may be contingent onthe occurrence of a specific event, such as activation of a signaltransduction pathway, and/or maybe manifest only in particular celltypes. The term “modulating 5-HT_(1A) receptors,” also refers toaltering the function of a 5-HT_(1A) receptor by increasing ordecreasing the probability that a complex forms between a 5-HT_(1A)receptor and a natural binding partner to form a multimer. A 5-HT_(1A)receptor modulator may increase the probability that such a complexforms between the 5-HT_(1A) receptor and the natural binding partner,may increase or decrease the probability that a complex forms betweenthe 5-HT_(2A) receptor and the natural binding partner depending on theconcentration of the compound exposed to the 5-HT_(1A) receptor, and ormay decrease the probability that a complex forms between the 5-HT_(1A)receptor and the natural binding partner. Furthermore, the term includesallosteric modulation of the receptor 5-HT_(1A), i.e., modulation of the5-HT_(1A) receptor through interaction with the 5-HT_(1A) receptor thatis topographically different than the orthosteric site recognized by thecell's endogenous agonist, such modulation further including positiveallosteric modulation (PAM), negative allosteric modulation (NAM) andsilent allosteric modulation (SAM).

The term “5-HT_(1A) receptor-mediated disorder”, as used herein, refersto a disorder that is characterized by abnormal 5-HT_(1A) receptoractivity. A 5-HT_(1A) receptor-mediated disorder may be completely orpartially mediated by modulating 5-HT_(2A) receptors. In particular, a5-HT_(1A) receptor-mediated disorder is one in which modulation of5-HT_(1A) receptors results in some effect on the underlying disordere.g., administration of a 5-HT_(1A) receptor modulator results in someimprovement in at least some of the subjects being treated.

The term “pharmaceutical formulation”, as used herein, refers to apreparation in a form which allows an active ingredient, including apsychoactive ingredient, contained therein to provide effectivetreatment, and which does not contain any other ingredients which causeexcessive toxicity, an allergic response, irritation, or other adverseresponse commensurate with a reasonable risk/benefit ratio. Thepharmaceutical formulation may contain other pharmaceutical ingredientssuch as excipients, carriers, diluents, or auxiliary agents.

The term “recreational drug formulation”, as used herein, refers to apreparation in a form which allows a psychoactive ingredient containedtherein to be effective for administration as a recreational drug, andwhich does not contain any other ingredients which cause excessivetoxicity, an allergic response, irritation, or other adverse responsecommensurate with a reasonable risk/benefit ratio. The recreational drugformulation may contain other ingredients such as excipients, carriers,diluents, or auxiliary agents.

The term “effective for administration as a recreational drug”, as usedherein, refers to a preparation in a form which allows a subject tovoluntarily induce a psychoactive effect for non-medical purposes uponadministration, generally in the form of self-administration. The effectmay include an altered state of consciousness, satisfaction, pleasure,euphoria, perceptual distortion, or hallucination.

The term “effective amount”, as used herein, refers to an amount of anactive agent, pharmaceutical formulation, or recreational drugformulation, sufficient to induce a desired biological or therapeuticeffect, including a prophylactic effect, and further including apsychoactive effect. Such effect can include an effect with respect tothe signs, symptoms or causes of a disorder, or disease or any otherdesired alteration of a biological system. The effective amount can varydepending, for example, on the health condition, injury stage, disorderstage, or disease stage, weight, or sex of a subject being treated,timing of the administration, manner of the administration, age of thesubject, and the like, all of which can be determined by those of skillin the art.

The terms “treating” and “treatment”, and the like, as used herein, areintended to mean obtaining a desirable physiological, pharmacological,or biological effect, and includes prophylactic and therapeutictreatment. The effect may result in the inhibition, attenuation,amelioration, or reversal of a sign, symptom or cause of a disorder, ordisease, attributable to the disorder, or disease, which includes mentaland psychiatric diseases and disorders. Clinical evidence of theprevention or treatment may vary with the disorder, or disease, thesubject, and the selected treatment.

The term “pharmaceutically acceptable”, as used herein, refers tomaterials, including excipients, carriers, diluents, or auxiliaryagents, that are compatible with other materials in a pharmaceutical orrecreational drug formulation and within the scope of reasonable medicaljudgement suitable for use in contact with a subject without excessivetoxicity, allergic response, irritation, or other adverse responsecommensurate with a reasonable risk/benefit ratio.

The term “psilocybin biosynthetic enzyme complement”, as used herein,refers to one or more polypeptides which alone or together are capableof facilitating the chemical conversion of a psilocybin precursorcompound and form another psilocybin precursor compound, or an aminatedpsilocybin derivative compound. A psilocybin biosynthetic enzymecomplement can include, for example, a tryptophan synthase subunit Bpolypeptide, a tryptophan decarboxylase and/or a N-acetyl transferase.

The term “tryptophan synthase subunit B polypeptide” as used herein,refers to any and all enzymes comprising a sequence of amino acidresidues which is (i) substantially identical to the amino acidsequences constituting any tryptophan synthase subunit B polypeptide setforth herein, including, for example, SEQ. ID NO: 9, or (ii) encoded bya nucleic acid sequence capable of hybridizing under at least moderatelystringent conditions to any nucleic acid sequence encoding anytryptophan synthase subunit B polypeptide set forth herein, but for theuse of synonymous codons.

The term “tryptophan decarboxylase” as used herein, refers to any andall enzymes comprising a sequence of amino acid residues which is (i)substantially identical to the amino acid sequences constituting anytryptophan decarboxylase polypeptide set forth herein, including, forexample, SEQ. ID NO: 12, or (ii) encoded by a nucleic acid sequencecapable of hybridizing under at least moderately stringent conditions toany nucleic acid sequence encoding any tryptophan decarboxylase setforth herein, but for the use of synonymous codons.

The term “N-acetyl transferase” as used herein, refers to any and allenzymes comprising a sequence of amino acid residues which is (i)substantially identical to the amino acid sequences constituting anyN-acetyl transferase polypeptide set forth herein, including, forexample, SEQ. ID NO: 5, or (ii) encoded by a nucleic acid sequencecapable of hybridizing under at least moderately stringent conditions toany nucleic acid sequence encoding any N-acetyl transferase set forthherein, but for the use of synonymous codons.

The term “N-methyl transferase” as used herein, refers to any and allenzymes comprising a sequence of amino acid residues which is (i)substantially identical to the amino acid sequences constituting anyN-methyl transferase polypeptide set forth herein, including, forexample, SEQ. ID NO: 14, or (ii) encoded by a nucleic acid sequencecapable of hybridizing under at least moderately stringent conditions toany nucleic acid sequence encoding any N-methyl transferase set forthherein, but for the use of synonymous codons.

The terms “nucleic acid sequence encoding tryptophan synthase subunit Bpolypeptide”, as may be used interchangeably herein, refer to any andall nucleic acid sequences encoding a tryptophan synthase subunit Bpolypeptide, including, for example, SEQ. ID NO: 8. Nucleic acidsequences encoding a tryptophan synthase subunit B polypeptide furtherinclude any and all nucleic acid sequences which (i) encode polypeptidesthat are substantially identical to the tryptophan synthase subunit Bpolypeptide sequences set forth herein; or (ii) hybridize to anytryptophan synthase subunit B polypeptide nucleic acid sequences setforth herein under at least moderately stringent hybridizationconditions or which would hybridize thereto under at least moderatelystringent conditions but for the use of synonymous codons.

The terms “nucleic acid sequence encoding tryptophan decarboxylase”, and“nucleic acid sequence encoding a tryptophan decarboxylase polypeptide”,as may be used interchangeably herein, refer to any and all nucleic acidsequences encoding a tryptophan decarboxylase, including, for example,SEQ. ID NO: 11. Nucleic acid sequences encoding a tryptophandecarboxylase polypeptide further include any and all nucleic acidsequences which (i) encode polypeptides that are substantially identicalto the tryptophan decarboxylase polypeptide sequences set forth herein;or (ii) hybridize to any tryptophan decarboxylase nucleic acid sequencesset forth herein under at least moderately stringent hybridizationconditions or which would hybridize thereto under at least moderatelystringent conditions but for the use of synonymous codons.

The terms “nucleic acid sequence encoding N-acetyl transferase”, and“nucleic acid sequence encoding an N-acetyl transferase polypeptide”, asmay be used interchangeably herein, refer to any and all nucleic acidsequences encoding an N-acetyl transferase, including, for example, SEQ.ID NO: 4. Nucleic acid sequences encoding an N-acetyl transferasepolypeptide further include any and all nucleic acid sequences which (i)encode polypeptides that are substantially identical to the N-acetyltransferase polypeptide sequences set forth herein; or (ii) hybridize toany N-acetyl transferase nucleic acid sequences set forth herein underat least moderately stringent hybridization conditions or which wouldhybridize thereto under at least moderately stringent conditions but forthe use of synonymous codons.

The terms “nucleic acid sequence encoding an N-methyl transferase”, and“nucleic acid sequence encoding an N-methyl transferase polypeptide”, asmay be used interchangeably herein, refer to any and all nucleic acidsequences encoding an N-methyl transferase, including, for example, SEQ.ID NO: 13. Nucleic acid sequences encoding an N-methyl transferasepolypeptide further include any and all nucleic acid sequences which (i)encode polypeptides that are substantially identical to the N-methyltransferase polypeptide sequences set forth herein; or (ii) hybridize toany N-methyl transferase nucleic acid sequences set forth herein underat least moderately stringent hybridization conditions or which wouldhybridize thereto under at least moderately stringent conditions but forthe use of synonymous codons.

The terms “nucleic acid”, or “nucleic acid sequence”, as used herein,refer to a sequence of nucleoside or nucleotide monomers, consisting ofnaturally occurring bases, sugars and intersugar (backbone) linkages.The term also includes modified or substituted sequences comprisingnon-naturally occurring monomers or portions thereof. The nucleic acidsof the present disclosure may be deoxyribonucleic nucleic acids (DNA) orribonucleic acids (RNA) and may include naturally occurring basesincluding adenine, guanine, cytosine, thymidine, and uracil. The nucleicacids may also contain modified bases. Examples of such modified basesinclude aza and deaza adenine, guanine, cytosine, thymidine and uracil,and xanthine and hypoxanthine. A sequence of nucleotide or nucleosidemonomers may be referred to as a polynucleotide sequence, nucleic acidsequence, a nucleotide sequence, or a nucleoside sequence.

The term “polypeptide”, as used herein in conjunction with a referenceSEQ. ID NO, refers to any and all polypeptides comprising a sequence ofamino acid residues which is (i) substantially identical to the aminoacid sequence constituting the polypeptide having such reference SEQ. IDNO, or (ii) encoded by a nucleic acid sequence capable of hybridizingunder at least moderately stringent conditions to any nucleic acidsequence encoding the polypeptide having such reference SEQ. ID NO, butfor the use of synonymous codons. A sequence of amino acid residues maybe referred to as an amino acid sequence, or polypeptide sequence.

The term “nucleic acid sequence encoding a polypeptide”, as used hereinin conjunction with a reference SEQ. ID NO, refers to any and allnucleic acid sequences encoding a polypeptide having such reference SEQ.ID NO. Nucleic acid sequences encoding a polypeptide, in conjunctionwith a reference SEQ. ID NO, further include any and all nucleic acidsequences which (i) encode polypeptides that are substantially identicalto the polypeptide having such reference SEQ. ID NO; or (ii) hybridizeto any nucleic acid sequences encoding polypeptides having suchreference SEQ. ID NO under at least moderately stringent hybridizationconditions or which would hybridize thereto under at least moderatelystringent conditions but for the use of synonymous codons.

By the term “substantially identical” it is meant that two amino acidsequences preferably are at least 70% identical, and more preferably areat least 85% identical and most preferably at least 95% identical, forexample 96%, 97%, 98% or 99% identical. In order to determine thepercentage of identity between two amino acid sequences the amino acidsequences of such two sequences are aligned, using for example thealignment method of Needleman and Wunsch (J. Mol. Biol., 1970, 48: 443),as revised by Smith and Waterman (Adv. Appl. Math., 1981, 2: 482) sothat the highest order match is obtained between the two sequences andthe number of identical amino acids is determined between the twosequences. Methods to calculate the percentage identity between twoamino acid sequences are generally art recognized and include, forexample, those described by Carillo and Lipton (SIAM J. Applied Math.,1988, 48:1073) and those described in Computational Molecular Biology,Lesk, e.d. Oxford University Press, New York, 1988, Biocomputing:Informatics and Genomics Projects. Generally, computer programs will beemployed for such calculations. Computer programs that may be used inthis regard include, but are not limited to, GCG (Devereux et al.,Nucleic Acids Res., 1984, 12: 387) BLASTP, BLASTN and FASTA (Altschul etal., J. Mol. Biol., 1990:215:403). A particularly preferred method fordetermining the percentage identity between two polypeptides involvesthe Clustal W algorithm (Thompson, J D, Higgines, D G and Gibson T J,1994, Nucleic Acid Res 22(22): 4673-4680 together with the BLOSUM 62scoring matrix (Henikoff S & Henikoff, J G, 1992, Proc. Natl. Acad. Sci.USA 89: 10915-10919 using a gap opening penalty of 10 and a gapextension penalty of 0.1, so that the highest order match obtainedbetween two sequences wherein at least 50% of the total length of one ofthe two sequences is involved in the alignment.

By “at least moderately stringent hybridization conditions” it is meantthat conditions are selected which promote selective hybridizationbetween two complementary nucleic acid molecules in solution.Hybridization may occur to all or a portion of a nucleic acid sequencemolecule. The hybridizing portion is typically at least 15 (e.g., 20,25, 30, 40 or 50) nucleotides in length. Those skilled in the art willrecognize that the stability of a nucleic acid duplex, or hybrids, isdetermined by the Tm, which in sodium containing buffers is a functionof the sodium ion concentration and temperature (Tm=81.5° C.-16.6 (Log10 [Na+])+0.41(% (G+C)−600/I), or similar equation). Accordingly, theparameters in the wash conditions that determine hybrid stability aresodium ion concentration and temperature. In order to identify moleculesthat are similar, but not identical, to a known nucleic acid molecule a1% mismatch may be assumed to result in about a 1° C. decrease in Tm,for example if nucleic acid molecules are sought that have a >95%identity, the final wash temperature will be reduced by about 5° C.Based on these considerations those skilled in the art will be able toreadily select appropriate hybridization conditions. In preferredembodiments, stringent hybridization conditions are selected. By way ofexample the following conditions may be employed to achieve stringenthybridization: hybridization at 5×sodium chloride/sodium citrate(SSC)/5×Denhardt's solution/1.0% SDS at Tm (based on the above equation)−5° C., followed by a wash of 0.2×SSC/0.1% SDS at 60° C. Moderatelystringent hybridization conditions include a washing step in 3×SSC at42° C. It is understood however that equivalent stringencies may beachieved using alternative buffers, salts, and temperatures. Additionalguidance regarding hybridization conditions may be found in: CurrentProtocols in Molecular Biology, John Wiley & Sons, N.Y., 1989,6.3.1.-6.3.6 and in: Sambrook et al., Molecular Cloning, a LaboratoryManual, Cold Spring Harbor Laboratory Press, 1989, Vol. 3.

The term “functional variant”, as used herein in reference topolynucleotides or polypeptides, refers to polynucleotides orpolypeptides capable of performing the same function as a notedreference polynucleotide or polypeptide. Thus, for example, a functionalvariant of the polypeptide set forth in SEQ. ID NO: 2, refers to apolypeptide capable of performing the same function as the polypeptideset forth in SEQ. ID NO: 2. Functional variants include modified apolypeptide wherein, relative to a noted reference polypeptide, themodification includes a substitution, deletion, or addition of one ormore amino acids. In some embodiments, substitutions are those thatresult in a replacement of one amino acid with an amino acid havingsimilar characteristics. Such substitutions include, without limitation(i) glutamic acid and aspartic acid; (i) alanine, serine, and threonine;(iii) isoleucine, leucine, and valine, (iv) asparagine and glutamine,and (v) tryptophan, tyrosine, and phenylalanine. Functional variantsfurther include polypeptides having retained or exhibiting an enhancedpsilocybin biosynthetic bioactivity.

The term “chimeric”, as used herein in the context of nucleic acids,refers to at least two linked nucleic acids which are not naturallylinked. Chimeric nucleic acids include linked nucleic acids of differentnatural origins. For example, a nucleic acid constituting a microbialpromoter linked to a nucleic acid encoding a plant polypeptide isconsidered chimeric. Chimeric nucleic acids also may comprise nucleicacids of the same natural origin, provided they are not naturallylinked. For example a nucleic acid constituting a promoter obtained froma particular cell-type may be linked to a nucleic acid encoding apolypeptide obtained from that same cell-type, but not normally linkedto the nucleic acid constituting the promoter. Chimeric nucleic acidsalso include nucleic acids comprising any naturally occurring nucleicacids linked to any non-naturally occurring nucleic acids.

The terms “substantially pure” and “isolated”, as may be usedinterchangeably herein describe a compound, e.g., a secondarymetabolite, psilocybin or a psilocybin derivative, polynucleotide, or apolypeptide, which has been separated from components that naturallyaccompany it. Typically, a compound is substantially pure when at least60%, more preferably at least 75%, more preferably at least 90%, 95%,96%, 97%, or 98%, and most preferably at least 99% of the total material(by volume, by wet or dry weight, or by mole percent or mole fraction)in a sample is the compound of interest. Purity can be measured by anyappropriate method, e.g., in the case of polypeptides, bychromatography, gel electrophoresis or HPLC analysis.

The term “recovered” as used herein in association with a chemicalcompound, refers to a more or less pure form of the chemical compound.

General Implementation

As hereinbefore mentioned, the present disclosure relates to psilocybinderivatives. In particular, the present disclosure provides novelaminated psilocybin derivatives. In general, the herein providedcompositions exhibit functional properties which deviate from thefunctional properties of psilocybin. Thus, for example, the aminatedpsilocybin derivatives, can exhibit pharmacological properties whichdeviate from psilocybin. Furthermore, the aminated derivatives maypsilocybin derivatives may exhibit physico-chemical properties whichdiffer from psilocybin. Thus, for example, aminated psilocybinderivatives may exhibit superior solubility in a solvent, for example,an aqueous solvent. The aminated psilocybin derivatives in this respectare useful in the formulation of pharmaceutical and recreational drugformulations. Furthermore, the aminated psilocybin compounds of thepresent disclosure may be used as a feedstock material for derivingfurther psilocybin derivatives. In one embodiment, the aminatedpsilocybin derivatives of the present disclosure can conveniently besynthetically produced. The practice of this method avoids theextraction of psilocybin from mushrooms and the performance ofsubsequent chemical reactions to achieve aminated derivatives.Furthermore, the growth of mushrooms can be avoided thus limiting thedependence on climate and weather, and potential legal and socialchallenges associated with the cultivation of mushrooms containingpsychoactive compounds. The method can efficiently yield substantialquantities of aminated psilocybin derivatives.

In what follows selected embodiments are described with reference to thedrawings.

Initially example aminated psilocybin derivatives will be described.Thereafter example methods of using and making the aminated psilocybinderivatives will be described.

Accordingly, in one aspect, the present disclosure provides derivativesof a compound known as psilocybin of which the chemical structure isshown in FIG. 1 . The derivatives herein provided are, in particular,derivatives of psilocybin including an amino group or N-substitutedamino group.

Thus, in one aspect, the present disclosure provides, in accordance withthe teachings herein, in at least one embodiment, a chemical compound orsalt thereof having formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group, O-alkyl group, ahydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen, an alkyl group, an aryl group or an acylgroup.

Thus, referring to the chemical compound having formula (I), initiallyit is noted that, in an aspect thereof, at least one of R₂, R₄, R₅, R₆,or R₇ is an amino group or N-substituted amino group.

Thus, referring to the chemical compound having the formula (I),initially it is noted that, in an aspect hereof, at least one of R₂, R₅,R₆, or R₇ is an amino group or an N-substituted amino group.

In a further aspect, at least one of R₂, R₅, R₆, or R₇ can be aN-substituted amino group, wherein one, or at least one, hydrogen atomis substituted by a group selected from an alkyl group, an aryl group,an acyl group, or a sulfonyl group. Thus, for example, in oneembodiment, the N-substituted amino group can be

a chemical group having the formula (XX):

wherein R′, and R″ are each independently selected from a hydrogen atom,an alkyl group, and an aryl group, provided however, that at least oneof R′ and R″ is not a hydrogen atom.

In a further example embodiment, the N-substituted amino group can be achemical group having the formula chemical group (XXI):

wherein R′, R″ and R′″ are each independently selected from a hydrogenatom, an alkyl group and an aryl group, provided however that at leastone of R′, R″, and R′″ is not a hydrogen atom.

In a further example embodiment, the N-substituted amino group can be achemical group having the formula (XXII) (an acyl group):

wherein R′, and R′″ are each independently selected from a hydrogenatom, an alkyl group, and an aryl group.

In a further example embodiment, the N-substituted amino group can be achemical group having the formula (XXIII) (a sulfonyl group):

wherein R′ and R″ are each independently selected from a hydrogen atom,an alkyl group and an aryl group.

In yet a further example embodiment, the N-substituted amino group canbe a chemical group having the formula (XXIV) (a sulfo group):

wherein R″ is selected from a hydrogen atom, an alkyl group, and an arylgroup. The nitrogen atom of chemical groups (XXII), (XXIII) and (XXIV)can also be positively charged and be further substituted with H, orR′″.

Continuing to refer to the chemical compound having formula (I), in afurther aspect, R₂, R₅, R₆, or R₇ can be a N-substituted amino-groupwherein two, or at least two, hydrogen atoms are substituted by a groupindependently selected from an alkyl group, an aryl group, an acyl groupor a sulfonyl group.

In at least one embodiment, at least one of R₂, R₅, R₆, or R₇ can be aN-substituted amino-group (i.e., an ammonium group), wherein threehydrogen atoms are substituted by a group independently selected from analkyl group, or an aryl group, wherein the nitrogen atom of theN-substituted group carries a positive charge.

Continuing to refer to the chemical compound having formula (I), in afurther aspect hereof, R_(3A) and R_(3B) can each independently be ahydrogen atom, an alkyl group, an acyl group or an aryl group. Thus,R_(3A) and R_(3B) can each be a hydrogen atom, or R_(3A) and R_(3B) caneach be an alkyl group, such as a methyl group, ethyl group, propylgroup, or longer chain alkyl group, or R_(3A) and R_(3B) can be each bean acyl group, or R_(3A) and R_(3B) can each be an aryl group.Furthermore, one of R_(3A) and R_(3B) can be a hydrogen atom, and one ofR_(3A) and R_(3B) can be an alkyl group. One of R_(3A) and R_(3B) can bea hydrogen atom, and one of R_(3A) and R_(3B) can be an acyl group. Oneof R_(3A) and R_(3B) can be a hydrogen atom, and one of R_(3A) andR_(3B) can be an aryl group. One of R_(3A) and R_(3B) can be an alkylgroup, and one of R_(3A) and R_(3B) can be an aryl group. One of R_(3A)and R_(3B) can be an alkyl group, and one of R_(3A) and R_(3B) can be anacyl group. One of R_(3A) and R_(3B) can be an acyl group, and one ofR_(3A) and R_(3B) can be an aryl group.

Continuing to refer to the chemical compound having formula (I), in afurther aspect hereof, R₄, when it is not an amino group orN-substituted amino group can be is a hydrogen atom, an alkyl group,O-alkyl group, a hydroxy group, or a phosphate group.

Continuing to refer to the chemical compound having formula (I), in afurther aspect hereof, the non-aminated groups R₂, R₅, R₆, or R₇ can bea hydrogen atom or an alkyl or O-alkyl group. Referring now to FIGS.3A-3D, examples of aminated psilocybin derivatives in accordanceherewith, wherein one of R₂, R₅, R₆, or R₇ are aminated, and thenon-aminated groups R₂, R₅, R₆, or R₇ are hydrogen atoms are: the2-amino-psilocybin derivative compound depicted in FIG. 3A, the5-amino-psilocybin derivative depicted in FIG. 3B, the6-amino-psilocybin derivative depicted in FIG. 3C, the7-amino-psilocybin derivative depicted in FIG. 3D,

Referring now to FIGS. 3E-3H, examples of aminated psilocybinderivatives in accordance herewith, wherein one of R₂, R₅, R₆, or R₇ areaminated, and the non-aminated groups R₂, R₅, R₆, or R₇ are hydrogenatoms are: the 2-N,N-substituted amino-psilocybin derivative compounddepicted in FIG. 3E, the 5-N,N-substituted amino-psilocybin derivativedepicted in FIG. 3F, the 6-N,N-substituted amino-psilocybin derivativedepicted in FIG. 3G, and the 7-N,N-substituted amino-psilocybinderivative depicted in FIG. 3H. It is noted that in FIGS. 3E-3H,R′_(2a), R″_(2b), R′_(5a), R″_(5b), R′_(6a), R″_(6b), R′_(7a), andR″_(7b) can each be independently selected from an alkyl group, an arylgroup, an acyl group, a sulfonyl group, a sulfo group, or a hydrogenatom, provided however, that N-substituted amino psilocybin derivativesdo not include psilocybin derivatives wherein in both R′_(2a), andR″_(2b), or both R′_(5a), and R″_(5b), or both R′_(6a) and R″_(6b), orboth R′_(7a), and R″_(7b) are hydrogen atoms.

In a further aspect hereof, the non-aminated groups R₂, R₅, R₆, or R₇can be a hydrogen atom or an alkyl or O-alkyl group. Referring now toFIGS. 3I-3L, examples of aminated psilocybin derivatives in accordanceherewith, wherein one of R₂, R₅, R₆, or R₇ are aminated, and thenon-aminated groups R₂, R₅, R₆, or R₇ are alkyl or O-alkyl groups are:the 2-amino-6-ethyl-psilocybin derivative compound depicted in FIG. 3I,the 5-amino-7-methyl psilocybin derivative depicted in FIG. 3J, the5-ethoxy-6-amino-psilocybin derivative depicted in FIG. 3K, the6-O-methyl-7-amino-psilocybin derivative depicted in FIG. 3L,

Referring now to FIGS. 3M-3P, examples of aminated psilocybinderivatives in accordance herewith, wherein one of R₂, R₅, R₆, or R₇ areaminated, and the non-aminated groups R₂, R₅, R₆, or R₇ are hydrogenatoms are: the 2-N,N-substituted amino-7methyl-psilocybin derivativecompound depicted in FIG. 3M, the 5-N,N-substitutedamino-6-ethyl-psilocybin derivative depicted in FIG. 3N, the5-O-methyl-6-N,N-substituted amino-psilocybin derivative depicted inFIG. 3O, and the 5-O-ethyl-7-N,N-substituted amino-psilocybin derivativedepicted in FIG. 3P. It is noted that in FIGS. 3M-3P, R′_(2a), R″_(2b),R′_(5a), R″_(5b), R′_(6a), R″_(6b), R′_(7a), and R″_(7b) can each beindependently selected from an alkyl group, an aryl group, an acylgroup, a sulfonyl group, a sulfo group, or a hydrogen atom, providedhowever, that N-substituted amino psilocybin derivatives do not includepsilocybin derivatives wherein in both R′_(2a), and R″_(2b), or bothR′_(5a), and R″_(5b), or both R′_(6a) and R″_(6b), or both R′_(7a), andR″_(7b) are hydrogen atoms.

Referring now to FIGS. 4A-4F, examples of aminated psilocybinderivatives in accordance herewith, wherein two of R₂, R₅, R₆, or R₇ areaminated, and the non-aminated groups R₂, R₅, R₆, or R₇ are hydrogenatoms are: the 2,5-di-amino-psilocybin derivative compound depicted inFIG. 4A, the 2,6-di-amino-psilocybin derivative depicted in FIG. 4B, the2,7-di-amino-psilocybin derivative depicted in FIG. 4C, the5,6-di-amino-psilocybin derivative depicted in FIG. 4D, the5,7-di-amino-psilocybin derivative depicted in FIG. 4E, and the6,7-di-amino-psilocybin derivative depicted in FIG. 4F.

Referring now to FIGS. 4G-4J, examples of aminated psilocybinderivatives in accordance herewith, wherein three of R₂, R₅, R₆, or R₇are aminated, and the non-aminated groups R₂, R₅, R₆, or R₇ are hydrogenatoms are: the 2,5,6-tri-amino-psilocybin derivative compound depictedin FIG. 4G, the 2,5,7-tri-amino-psilocybin derivative depicted in FIG.4H, the 2,6,7-tri-amino-psilocybin derivative depicted in FIG. 4I, andthe 5,6,7-tri-amino-psilocybin derivative depicted in FIG. 4J.

Referring now to FIG. 4K an example of a aminated psilocybin derivativesin accordance herewith, wherein all four of R₂, R₅, R₆, or R₇ areaminated is the 2,5,6,7-tetra-amino-psilocybin derivative depicted inFIG. 4K.

Referring now to FIGS. 4L-4Q, examples of aminated psilocybinderivatives in accordance herewith, wherein two of R₂, R₅, R₆, or R₇ areaminated, and the non-aminated groups R₂, R₅, R₆, or R₇ are hydrogenatoms are: the 2-N,5-N,N-di-substituted-amino-psilocybin derivativecompound depicted in FIG. 4L, the2-N,6-N,N-di-substituted-amino-psilocybin derivative depicted in FIG.4M, the 2-N,7-N,N-di-substituted-amino-psilocybin derivative depicted inFIG. 4N, the 5-N,6-N,N-di-substituted-amino-psilocybin derivativedepicted in FIG. 4O, the 5-N,7-N,N-di-substituted-amino-psilocybinderivative depicted in FIG. 4P, and the6-N,N-7-N,N-di-substituted-amino-psilocybin derivative depicted in FIG.4Q. As hereinbefore noted, the substituents may be a group selected froman alkyl group, an aryl group, an acyl group, or a sulfonyl group. Thus,by way of example only, in the 5-N,6-N,N-di-substituted-amino-psilocybinderivative depicted in FIG. 4O at least one of R′_(5a), of R″_(5b), andat least one of R′_(6a), and R″_(6b), is an alkyl group, an aryl group,an acyl group, a sulfo group, or a sulfonyl group.

Continuing to refer to FIGS. 4L-4Q, it is noted that, in otherembodiments, instead of being di-substituted, only one of the aminatedgroups may be an N-substituted amino group while the other aminatedgroup is an amino group. Thus, for example, referring to FIG. 4P, insuch embodiments, only R″_(7a) or R′_(7b) may be N-substituted, whileR′_(5a) and R″_(5b) may each be a hydrogen atom, thus forming an aminogroup, or conversely, only R″_(5a) or R′_(5b) may be N-substituted,while R′_(7a) and R″_(7b) may be hydrogen atoms thus forming an aminogroup. It is to be understood that any and all embodiments including theaminated psilocybin derivatives shown in FIGS. 4L-4Q, provided that atleast one of the R′_(2a), R″_(2b), R′_(5a), R″_(5b), R′_(6a), R″_(6b),R′_(7a), and R″_(7b) is an N-substituted amino group are also includedherein. As hereinbefore noted, the substituents may be a group selectedfrom an alkyl group, an aryl group, an acyl group, a sulfo group, or asulfonyl group.

Referring now to FIGS. 4R-4U, examples of aminated psilocybinderivatives in accordance herewith, wherein three of R₂, R₅, R₆, or R₇are aminated, and the non-aminated groups R₂, R₅, R₆, or R₇ are hydrogenatoms are: the 2-N,5-N,6-N,N-tri-substituted-amino-psilocybin derivativecompound depicted in FIG. 4R, the2-N,5-N,7-N,N-tri-substituted-amino-psilocybin derivative depicted inFIG. 4S, the 2-N,6-N,7-N,N-tri-substituted amino-psilocybin derivativedepicted in FIG. 4T, and the5-N,6-N,7-N,N-tri-substituted-amino-psilocybin derivative depicted inFIG. 4U. As hereinbefore noted, the substituents may be a group selectedfrom an alkyl group, an aryl group, an acyl group, or a sulfonyl group.Thus, by way of example only, in the5-N,6-N,7-N,N-tri-substituted-amino-psilocybin derivative depicted inFIG. 4U at least one of R′_(5a), of R″_(5b), and at least one ofR′_(6a), and R″_(6b), and at least one of R′_(7a), and R″_(7b) is analkyl group, an aryl group, an acyl group, a sulfo group, or a sulfonylgroup.

Continuing to refer to FIGS. 4R-4U, it is noted that, in otherembodiments, instead of being tri-substituted, only one or two of theaminated groups may be an N-substituted amino group while the otheraminated groups is/are an amino group. Thus, for example, referring toFIG. 4S, in such embodiments, only R″_(7a) or R′_(7b) may beN-substituted, while R′_(5a) and R″_(5b) and R′_(2a) and R″_(2b) may allbe a hydrogen atoms, thus forming two an amino groups, or R″_(5a) orR′_(5b) may be N-substituted, and R′_(7a) and R″_(7b) may N-substituted,but R′_(2a) and R″_(2b) may be hydrogen atoms thus forming an aminogroup. It is to be understood that any and all embodiments includingaminated psilocybin derivatives shown in FIGS. 4R-4U, provided that atleast one of the R′_(2a), R″_(2b), R′_(5a), R″_(5b), R′_(6a), R″_(6b),R′_(7a), and R″_(7b) is an N-substituted amino group are also includedherein. As hereinbefore noted, the substituents may be a group selectedfrom an alkyl group, an aryl group, an acyl group, a sulfo group, or asulfonyl group.

Referring now to FIG. 4V, an example of an aminated psilocybinderivatives in accordance herewith, wherein all four of R₂, R₅, R₆, orR₇ are aminated is the2-N,5-N,6-N,7-N,N-tetra-substituted-amino-psilocybin derivative depictedin FIG. 4K. As hereinbefore noted, the substituents may be a groupselected from an alkyl group, an aryl group, an acyl group, or asulfonyl group. Thus, by way of example only, in the2-N,N-5-N,6-N,7-N,N-tri-substituted-amino-psilocybin derivative depictedin FIG. 4V at least one of R′_(2a), of R″_(2b) and R′_(5a), of R″_(5b),at least one of R′_(6a), and R″_(6b), at least one of R′_(7a), andR″_(7b) is an alkyl group, an aryl group, an acyl group, a sulfo group,or a sulfonyl group.

Continuing to refer to FIG. 4V, it is noted that, in other embodiments,instead of being tetra-substituted, only one, two or three of theaminated groups may be an N-substituted amino group while the otheraminated groups is/are an amino group. Thus, for example, referring toFIG. 4V, in such embodiments, only R″_(7a) or R′_(7b) may beN-substituted, while R′_(5a) and R″_(5b), R′_(6a) and R″_(6b) andR′_(2a) and R″_(2b) may all be a hydrogen atoms, thus forming threeamino groups, or, for example R″_(5a) or R′_(5b) may be N-substituted,and R′_(7a) and R″_(7b) may N-substituted, but R′_(2a) and R″_(2b) andR′_(6a) and R″_(6b) may be hydrogen atoms thus forming two amino groups.It is to be understood that any and all embodiments including theaminated psilocybin derivatives shown in FIGS. 4V, provided that atleast one of the R′_(2a), R″_(2b), R′_(5a), R″_(5b), R′_(6a), R″_(6b),R′_(7a), and R″_(7b) is an N-substituted amino group are also includedherein. As hereinbefore noted, the substituents may be a group selectedfrom an alkyl group, an aryl group, an acyl group, a sulfo group, or asulfonyl group.

In a further aspect, R₄, can be an O-alkyl group. Referring now to FIGS.5A, 5B, 6A, 6B, 7A, and 7B examples of aminated psilocybin derivativesin accordance herewith, wherein R₅, and/or R₇ are amino groups and R₄ isan O-alkyl group are: the 4-O-methyl-5-amino-psilocybin derivativedepicted in FIG. 5A, the 4-O-ethyl-5-amino-psilocybin derivativedepicted in FIG. 5B, the 4-O-methyl-7-amino-psilocybin derivativedepicted in FIG. 6A, the 4-O-ethyl-7-amino-psilocybin derivativedepicted in FIG. GB, the 4-O-methyl-5,7-di-amino-psilocybin derivativedepicted in FIG. 7A, the 4-O-ethyl-5,7-di-amino-psilocybin derivativedepicted in FIG. 7B. It is noted that in these specific examples only5-amino, 7-amino, and 5,7-di-O-alkyl psilocybin derivatives are shown.Further examples of O-alkyl psilocybin derivatives included herein areany and all O-alkyl psilocybin derivatives which may be selected byreferring to the chemical formulas shown in FIGS. 4A-4J, wherein R₄ isan O-alkyl group. It will thus be clearly understood that FIGS. 5A, 5B,6A, 6B, 7A, and 7B represent examples only of aminated psilocybinderivatives having chemical formula (I) wherein non-aminated groups R₂,R₅, R₆, or R₇ are a hydrogen atom. Other aminated psilocybin derivativeswherein non-aminated groups R₂, R₅, R₆, or R₇ are a hydrogen atom canreadily be selected, and thus are included in the O-alkylated aminatedpsilocybin derivatives compounds of the present disclosure.

It is noted that the example aminated psilocybin derivatives shown inFIGS. 5A, 5B, 6A, 6B, 7A, and 7B are aminated psilocybin derivativescompounds by virtue of their amino groups. Considering FIGS. 5A, 5B, 6A,6B, 7A, and 7B, in conjunction with FIGS. 4L, 4N, and 4P, it is noted,and it will be clear that, in other embodiments, included herein are,further, aminated psilocybin derivatives FIGS. 5A, 5B, 6A, 6B, 7A, and7B, wherein instead of an amino group the psilocybin derivativepossesses at least on N-substituted amino group, i.e., R′_(7a) orR″_(7b) is substituted by a group selected from an alkyl group, an arylgroup, an acyl group, a sulfo group, or a sulfonyl group.

In a further aspect, R₄, can be an O-acyl group. Referring now to FIGS.5C, 5D, 6C, 6D, 7C, and 7D examples of aminated psilocybin derivativesin accordance herewith, wherein R₅, and/or R₇ are amino groups and R₄ isan O-acyl group are: the 4-acetyl-5-amino-psilocybin derivative depictedin FIG. 5C, the 4-propanoyl-5-amino-psilocybin derivative depicted inFIG. 5D, the 4-acetyl-7-amino-psilocybin derivative depicted in FIG. GC,the 4-propanoyl-7-amino-psilocybin derivative depicted in FIG. 6D, the4-acetyl-5,7-di-amino-psilocybin derivative depicted in FIG. 7C, the4-propanoyl-5,7-di-amino-psilocybin derivative depicted in FIG. 7D. Itis noted that in these specific examples only 5-amino, 7-amino, and5,7-di-O-acyl psilocybin derivatives are shown. Further examples ofO-acyl psilocybin derivatives included herein are any and all O-acylpsilocybin derivatives which may be selected by referring to thechemical formulas shown in FIGS. 4A-4J, wherein R₄ is an O-acyl group.It will thus be clearly understood that FIGS. 5C, 5D, 6C, 6D, 7C, and 7Drepresent examples only of O-acylated psilocybin derivatives havingchemical formula (I) wherein non-aminated groups R₂, R₅, R₆, or R₇ are ahydrogen atom. Other aminated psilocybin derivatives whereinnon-aminated groups R₂, R₅, R₆, or R₇ are a hydrogen atom can readily beselected, and thus are included in the aminated O-acylated psilocybinderivatives compounds of the present disclosure.

It is noted that the example aminated psilocybin derivatives shown inFIGS. 5C, 5D, 6C, 6D, 7C, and 7D are aminated psilocybin derivativescompounds by virtue of their amino groups. Considering 5C, 5D, 6C, 6D,7C, and 7D, in conjunction with FIGS. 4L, 4N, and 4P, it is noted, andit will be clear that, in other embodiments, included herein are,further, aminated psilocybin derivatives 5C, 5D, 6C, 6D, 7C, and 7D,wherein instead of an amino group the psilocybin derivative possesses atleast on N-substituted amino group, i.e., R′7a or R″_(7b) is substitutedby a group selected from an alkyl group, an aryl group, an acyl group, asulfo group, or a sulfonyl group.

In a further aspect, R₄ can be a hydroxy group. Referring now to FIGS.5E, 6E, and 7E examples of aminated psilocybin derivatives in accordanceherewith, wherein R₅, and/or R₇ are amino groups and are R₄ is a hydroxygroup are: the 4-hydroxy-5-amino-psilocybin derivative depicted in FIG.5E, the 4-hydroxy-7-amino-psilocybin derivative depicted in FIG. 6E, andthe 4-hydroxy-5,7-di-amino-psilocybin derivative depicted in FIG. 7E, Itis noted that in these specific examples only 5-amino, 7-amino, and5,7-di-hydroxy-psilocybin derivatives are shown. Further examples ofhydroxy-psilocybin derivatives included herein are any and allhydroxy-psilocybin derivatives which may be selected by referring to thechemical formulas shown in FIGS. 4A-4J, wherein R₄ is a hydroxy group.It will thus be clearly understood that FIGS. 5E, 6E, and 7E representexamples only of hydroxy psilocybin derivatives having chemical formula(I) wherein non-aminated groups R₂, R₅, R₆, or R₇ are a hydrogen atom.Other aminated psilocybin derivatives wherein non-aminated groups R₂,R₅, R₆, or R₇ are a hydrogen atom can readily be selected, and thus areincluded in the aminated hydroxy psilocybin derivatives compounds of thepresent disclosure.

It is noted that the example aminated psilocybin derivatives shown inFIGS. 5E, 6E, and 7E are aminated psilocybin derivatives compounds byvirtue of their amino groups. Considering FIGS. 5E, 6E, and 7E, inconjunction with FIGS. 4L, 4N, and 4P, it is noted, and it will be clearthat, in other embodiments, included herein are, further, aminatedpsilocybin derivatives FIGS. 5E, 6E, and 7E, wherein instead of an aminogroup the psilocybin derivative possesses at least on N-substitutedamino group, i.e., R′_(7a) or R″_(7b) is substituted by a group selectedfrom an alkyl group, an aryl group, an acyl group, a sulfo group, or asulfonyl group.

In a further aspect, R₄ can be a phosphate group. Referring now to FIGS.5F, 6F, and 7F examples of aminated psilocybin derivatives in accordanceherewith, wherein R₅, and/or R₇ are amino groups and are R₄ is aphosphate group are: the 4-phospho-5-amino-psilocybin derivativedepicted in FIG. 5F, the 4-phospho-7-amino-psilocybin derivativedepicted in FIG. 6F, and the 4-phosphate-5,7-amino-psilocybin derivativedepicted in FIG. 7F, It is noted that in these specific examples only5-amino, 7-amino, and 5,7-di-phospho-psilocybin derivatives are shown.Further examples of phosphate-psilocybin derivatives included herein areany and all phosphate-psilocybin derivatives which may be selected byreferring to the chemical formulas shown in FIGS. 4A-4J, wherein R₄ is aphosphate group. It will thus be clearly understood that FIGS. 5F, 6F,and 7F represent examples only of phosphate psilocybin derivativeshaving chemical formula (I) wherein non-aminated groups R₂, R₅, R₆, orR₇ are a hydrogen atom. Other aminated psilocybin derivatives whereinnon-aminated groups R₂, R₅, R₆, or R₇ are a hydrogen atom can readily beselected, and thus are included in the aminated phosphate psilocybinderivatives compounds of the present disclosure.

It is noted that the example aminated psilocybin derivatives shown inFIGS. 5F, 6F, and 7F are aminated psilocybin derivatives compounds byvirtue of their amino groups. Considering FIGS. 5F, 6F, and 7F, inconjunction with FIGS. 4L, 4N, and 4P, it is noted, and it will be clearthat, in other embodiments, included herein are, further, aminatedpsilocybin derivatives FIGS. 5F, 6F, and 7F, wherein instead of an aminogroup the psilocybin derivative possesses at least on N-substitutedamino group, i.e., R′_(7a) or R″_(7b) is substituted by a group selectedfrom an alkyl group, an aryl group, an acyl group, a sulfo group, or asulfonyl group.

In a further aspect, R₄ can be a hydrogen atom. Referring now to FIGS.5G, 6G, and 7G examples of aminated psilocybin derivatives in accordanceherewith, wherein R₅, and/or R₇ are amino groups and are R₄ is ahydrogen atom are: the 5-amino-psilocybin derivative depicted in FIG.5G, the 7-amino-psilocybin derivative depicted in FIG. 6G, and the5,7-di-amino-psilocybin derivative depicted in FIG. 7G, It is noted thatin these specific examples only 5-amino, 7-amino, and5,7-di-amino-hydro-psilocybin derivatives are shown. Further examples ofhydro-psilocybin derivatives included herein are any and allhydro-psilocybin derivatives which may be selected by referring to thechemical formulas shown in FIGS. 4A-4J, wherein R₄ is a hydrogen atom.It will thus be clearly understood that FIGS. 5G, 6G, and 7G representexamples only of hydro psilocybin derivatives having chemical formula(I) wherein non-aminated groups R₂, R₅, R₆, or R₇ are a hydrogen atom.Other aminated psilocybin derivatives wherein non-aminated groups R₂,R₅, R₆, or R₇ are a hydrogen atom can readily be selected, and thus areincluded in the aminated hydro psilocybin derivatives compounds of thepresent disclosure.

It is noted that the example aminated psilocybin derivatives shown inFIGS. 5G, 6G, and 7G are aminated psilocybin derivatives compounds byvirtue of their amino groups. Considering FIGS. 5G, 6G, and 7G, inconjunction with FIGS. 4L, 4N, and 4P, it is noted, and it will be clearthat, in other embodiments, included herein are, further, aminatedpsilocybin derivatives FIGS. 5G, 6G, and 7G, wherein instead of an aminogroup the psilocybin derivative possesses at least one N-substitutedamino group, i.e., R′_(7a) or R″_(7b) is substituted by a group selectedfrom an alkyl group, an aryl group, an acyl group, a sulfo group, or asulfonyl group.

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (III):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (IV):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (V):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (VI):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (VII):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (VIII):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (IX):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (X):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XI):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XII):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XIII):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XIV):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XV):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XVI):

Furthermore, in one embodiment, an aminated psilocybin derivativeaccording to the present disclosure can be a chemical compound havingthe formula (XVII):

Furthermore, it is noted that the aminated psilocybin derivatives of thepresent disclosure include salts thereof, including pharmaceuticallyacceptable salts. Thus, the nitrogen atom of the ethyl-amino groupextending in turn from the C₃ atom may be protonated, and the positivecharge may be balanced by, for example, chloride or sulfate ions, tothereby form a chloride salt or a sulfate salt. Furthermore, incompounds wherein R₄ is a phosphate group, the phosphate group may bede-protonated, and the negative charge may be balanced by, for example,sodium ions or potassium ions, to thereby form a sodium salt or apotassium salt.

Furthermore, it is noted that when R₄ is a phosphate group, the termaminated psilocybin derivative also includes compounds having theformula (XVIII):

wherein at least one of R₂, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein any R₂, R₅, R₆, or R₇ which arenot an amino group or N-substituted amino group are a hydrogen atom, analkyl group or O-alkyl group, and wherein R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, and aryl group or an acylgroup. Further included are salts of aminated psilocybins having theformula (XVIII), such as a sodium salt, a potassium salt etc.

Thus, to briefly recap, the present disclosure provides aminatedpsilocybin derivatives. The disclosure provides, in particular, achemical compound or salt thereof having formula (I):

wherein in an aspect, at least one of R₂, R₄, R₅, R₆, or R₇ is an aminogroup or N-substituted amino group. In an aspect, in formula (I), eachnon-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkyl group orO-alkyl group. In a further aspect, in formula (I), R₄ when it is notaminated is a hydrogen atom, an alkyl group or O-alkyl group, a hydroxygroup, or a phosphate group. Yet in a further aspect, R_(3A) and R_(3B)are each independently a hydrogen atom, an alkyl group, an aryl group,or an acyl group.

In one embodiment of the disclosure, a chemical compound or salt thereofhaving formula (I) is included:

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, alkyl group or O-alkyl group, ahydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group.

In one embodiment, at least one of R₂, R₄, R₅, R₆, or R₇ is an aminogroup or N-substituted amino group, and wherein each non-aminated R₂,R₅, R₆, or R₇ is a hydrogen atom or a (C₁-C₂₀)-alkyl group or(C₁-C₂₀)—O-alkyl group. In another embodiment, each non-aminated R₂, R₅,R₆, or R₇ is a hydrogen atom, a methyl group, ethyl group, a propylgroup, an O-methyl group, an O-ethyl group, or an O-propyl group.

In another embodiment, each non-aminated R₂, R₅, R₆, or R₇ is a hydrogenatom or a (C₁-C₁₀)-alkyl group or (C₁-C₁₀)—O-alkyl group. In anotherembodiment, each non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, amethyl group, ethyl group, a propyl group, an O-methyl group, an O-ethylgroup, or an O-propyl group.

In another embodiment, each non-aminated R₂, R₅, R₆, or R₇ is a hydrogenatom or a (C₁-C₆)-alkyl group or (C₁-C₆)—O-alkyl group. In anotherembodiment, each non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, amethyl group, ethyl group, a propyl group, an O-methyl group, an O-ethylgroup, or an O-propyl group.

In another embodiment, when R₄ is not aminated, R₄ is a hydrogen atom, a(C₁-C₂₀)-alkyl group or (C₁-C₂₀)—O-alkyl group, a hydroxy group, or aphosphate group. In another embodiment, when R₄ is not aminated, R₄ is ahydrogen atom, a (C₁-C₁₀)-alkyl group or (C₁-C₁₀)—O-alkyl group, ahydroxy group, or a phosphate group. In another embodiment, when R₄ isnot aminated, R₄ is a hydrogen atom, a (C₁-C₆)-alkyl group or(C₁-C₆)—O-alkyl group, a hydroxy group, or a phosphate group. In anotherembodiment, when R₄ is not aminated, R₄ is a hydrogen atom, a methylgroup, an ethyl group, a propyl group, a phosphate group, an O-methylgroup, an O-ethyl group, or an O-propyl group.

In another embodiment, R_(3A) and R_(3B) are each independently ahydrogen atom, a (C₁-C₂₀)-alkyl group, a (C₆-C₁₄)-aryl group, or a—C(═O)(C₁-C₂₀)-alkyl group. In another embodiment, R_(3A) and R_(3B) areeach independently a hydrogen atom, a (C₁-C₁₀)-alkyl group, a(C₆-C₁₀)-aryl group, or a —C(═O)(C₁-C₁₀)-alkyl group. In anotherembodiment, R_(3A) and R_(3B) are each independently a hydrogen atom, a(C₁-C₆)-alkyl group, a phenyl group, or a —C(═O)(C₁-C₆)-alkyl group. Inanother embodiment, R_(3A) and R_(3B) are each independently a hydrogenatom, a methyl group, an ethyl group, a propyl group, a phenyl group,—C(═O)—CH₃, —C(═O)—CH₂CH₃, or —C(═O)—CH₂CH₂CH₃.

In one embodiment of the disclosure, a chemical compound or salt thereofhaving formula (I) is included:

wherein R₂, R₅, R₆, and R₇ are independently or simultaneously ahydrogen atom, an alkyl group or O-alkyl group or an amino group orN-substituted amino group, R_(3A) and R_(3B) are each independently ahydrogen atom, an alkyl group, an aryl group, or an acyl group; and R₄is hydrogen atom, alkyl group or O-alkyl group, an amino group orN-substituted amino group, a hydroxy group, or a phosphate group;wherein at least one of R₂, R₄ R₅, R₆, and R₇ is an amino group orN-substituted amino group.

In one embodiment, R₂, R₅, R₆, and R₇ are independently orsimultaneously a hydrogen atom, (C₁-C₂₀)-alkyl group or (C₁-C₂₀)—O-alkylgroup or an amino group or N-substituted amino group. In one embodiment,R₂, R₅, R₆, and R₇ are independently or simultaneously a hydrogen atom,(C₁-C₁₀)-alkyl group or (C₁-C₁₀)—O-alkyl group or an amino group orN-substituted amino group. In one embodiment, R₂, R₅, R₆, and R₇ areindependently or simultaneously a hydrogen atom, (C₁-C₆)-alkyl group or(C₁-C₆)—O-alkyl group or an amino group or N-substituted amino group. Inone embodiment, R₂, R₅, R₆, and R₇ are independently or simultaneously ahydrogen atom, methyl, ethyl, propyl, O-methyl, O-ethyl, O-propyl, or anamino group or N-substituted amino group.

In one embodiment, R₄ is a hydrogen atom, (C₁-C₂₀)-alkyl group or(C₁-C₂₀)—O-alkyl group, an amino group or N-substituted amino group or aphosphate group. In one embodiment, R₄ is a hydrogen atom,(C₁-C₁₀)-alkyl group or (C₁-C₁₀)—O-alkyl group, an amino group orN-substituted amino group or a phosphate group. In one embodiment, R₄ isa hydrogen atom, (C₁-C₆)-alkyl group or (C₁-C₆)—O-alkyl group, an aminogroup or N-substituted amino group, a hydroxy group, or a phosphategroup. In one embodiment, R₄ is a hydrogen atom, methyl, ethyl, propyl,O-methyl, O-ethyl, O-propyl, an amino group or N-substituted aminogroup, a hydroxy group, or a phosphate group.

In another embodiment, R_(3A) and R_(3B) are each independently ahydrogen atom, a (C₁-C₂₀)-alkyl group, a (C₆-C₁₄)-aryl group, or a—C(═O)(C₁-C₂₀)-alkyl group. In another embodiment, R_(3A) and R_(3B) areeach independently a hydrogen atom, a (C₁-C₁₀)-alkyl group, a(C₆-C₁₀)-aryl group, or a —C(═O)(C₁-C₁₀)-alkyl group or O-alkyl group.In another embodiment, R_(3A) and R_(3B) are each independently ahydrogen atom, a (C₁-C₆)-alkyl group, a phenyl group, or a—C(═O)(C₁-C₆)-alkyl group. In another embodiment, R_(3A) and R_(3B) area hydrogen atom, a methyl group, an ethyl group, a propyl group, aphenyl group, —C(═O)—CH₃, —C(═O)—CH₂CH₃, or —C(═O)—CH₂CH₂CH₃.

The aminated psilocybin derivatives of the present disclosure may beused to prepare a pharmaceutical or recreational drug formulation. Thusin one embodiment, the present disclosure further provides in anotheraspect, pharmaceutical and recreational drug formulations comprisingaminated psilocybin derivatives. Accordingly, in one aspect, the presentdisclosure provides in a further embodiment a pharmaceutical orrecreational drug formulation comprising a chemical compound havingformula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom an alkyl group, an aryl group, or anacyl group, or a slat of the chemical compound, together with a diluent,carrier or excipient.

The pharmaceutical or recreational drug formulations may be prepared asliquids, tablets, capsules, microcapsules, nanocapsules, trans-dermalpatches, gels, foams, oils, aerosols, nanoparticulates, powders, creams,emulsions, micellar systems, films, sprays, ovules, infusions, teas,decoctions, suppositories, etc. and include a pharmaceuticallyacceptable salt or solvate of the aminated psilocybin compound togetherwith an excipient. The term “excipient” as used herein means anyingredient other than the chemical compound of the disclosure. As willreadily be appreciated by those of skill in art, the selection ofexcipient may depend on factors such as the particular mode ofadministration, the effect of the excipient on solubility of thechemical compounds of the present disclosure and methods for theirpreparation will be readily apparent to those skilled in the art. Suchcompositions and methods for their preparation may be found, forexample, in “Remington's Pharmaceutical Sciences”, 22^(nd) Edition(Pharmaceutical Press and Philadelphia College of Pharmacy at theUniversity of the Sciences, 2012).

The dose when using the compounds of the present disclosure can varywithin wide limits, and as is customary and is known to those of skillin the art, the dose can be tailored to the individual conditions ineach individual case. The dose depends, for example, on the nature andseverity of the illness to be treated, on the condition of the patient,on the compound employed or on whether an acute or chronic disease stateis treated, or prophylaxis is conducted, on the mode of delivery of thecompound, or on whether further active compounds are administered inaddition to the compounds of the present disclosure. Representativedoses of the present disclosure include, but are not limited to, about0.001 mg to about 5000 mg, about 0.001 mg to about 2500 mg, about 0.001mg to about 1000 mg, about 0.001 mg to about 500 mg, about 0.001 mg toabout 250 mg, about 0.001 mg to about 100 mg, about 0.001 mg to about 50mg, and about 0.001 mg to about 25 mg. Representative doses of thepresent disclosure include, but are not limited to, about 0.0001 toabout 1,000 mg, about 10 to about 160 mg, about 10 mg, about 20 mg,about 40 mg, about 80 mg, or about 160 mg. Multiple doses may beadministered during the day, especially when relatively large amountsare deemed to be needed, for example 2, 3 or 4, doses. Depending on thesubject and as deemed appropriate from the patient's physician or caregiver it may be necessary to deviate upward or downward from the dosesdescribed herein.

The pharmaceutical and drug formulations comprising the aminatedpsilocybin derivatives of the present disclosure may be administeredorally. Oral administration may involve swallowing, so that the compoundenters the gastrointestinal tract, or buccal or sublingualadministration may be employed by which the compound enters the bloodstream directly from the mouth. Formulations suitable for oraladministration include both solid and liquid formulations.

Solid formulations include tablets, capsules (containing particulates,liquids, microcapsules, or powders), lozenges (including liquid-filledlozenges), chews, multi- and nano-particulates, gels, solid solutions,liposomal preparations, microencapsulated preparations, creams, films,ovules, suppositories, and sprays.

Liquid formulations include suspensions, solutions, syrups, and elixirs.Such formulations may be employed as fillers in soft or hard capsulesand typically comprise a carrier, for example, water, ethanol,polyethylene glycol, propylene glycol, methylcellulose, or a suitableoil, and one or more emulsifying agents and/or suspending agents. Liquidformulations may also be prepared by the reconstitution of a solid, forexample, from a sachet.

Binders are generally used to impart cohesive qualities to a tabletformulation. Suitable binders include microcrystalline cellulose,gelatin, sugars, polyethylene glycol, natural and synthetic gums,polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose andhydroxypropyl methylcellulose.

Tablets may also contain diluents, such as lactose (monohydrate,spray-dried monohydrate, anhydrous and the like), mannitol, xylitol,dextrose, sucrose, sorbitol, microcrystalline cellulose, starch, anddibasic calcium phosphate dihydrate.

Tablets may also optionally comprise surface active agents, such assodium lauryl sulfate and polysorbate 80. When present, surface activeagents may comprise from 0.2% (w/w) to 5% (w/w) of the tablet.

Tablets may further contain lubricants such as magnesium stearate,calcium stearate, zinc stearate, sodium stearyl fumarate, and mixturesof magnesium stearate with sodium lauryl sulphate. Lubricants generallycomprise from 0.25% (w/w) to 10% (w/w), from 0.5% (w/w) to 3% (w/w) ofthe tablet.

In addition to the aminated psilocybin derivative, tablets may contain adisintegrant. Examples of disintegrants include sodium starch glycolate,sodium carboxymethyl cellulose, calcium carboxymethyl cellulose,croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substitutedhydroxypropyl cellulose, starch, pregelatinized starch and sodiumalginate. Generally, the disintegrant will comprise from 1% (w/w) to 25%(w/w) or from 5% (w/w) to 20% (w/w) of the dosage form.

Other possible auxiliary ingredients include anti-oxidants, colourants,flavouring agents, preservatives, and taste-masking agents.

For tablet dosage forms, depending on the desired effective amount ofthe chemical compound, the chemical compound of the present disclosuremay make up from 1% (w/w) to 80% (w/w) of the dosage form, moretypically from 5% (w/w) to 60% (w/w) of the dosage form.

Exemplary tablets contain up to about 80% (w/w) of the chemicalcompound, from about 10% (w/w) to about 90% (w/w) binder, from about 0%(w/w) to about 85% (w/w) diluent, from about 2% (w/w) to about 10% (w/w)disintegrant, and from about 0.25% (w/w) to about 10% (w/w) lubricant.

The formulation of tablets is discussed in “Pharmaceutical Dosage Forms:Tablets”, Vol. 1-Vol. 3, by CRC Press (2008).

The pharmaceutical and recreational drug formulations comprising theaminated psilocybin derivatives of the present disclosure may also beadministered directly into the blood stream, into muscle, or into aninternal organ. Thus, the pharmaceutical and recreational drugformulations can be administered parenterally (for example, bysubcutaneous, intravenous, intraarterial, intrathecal, intraventricular,intracranial, intramuscular, or intraperitoneal injection). Parenteralformulations are typically aqueous solutions which may containexcipients such as salts, carbohydrates and buffering agents (in oneembodiment, to a pH of from 3 to 9), but, for some applications, theymay be more suitably formulated as a sterile non-aqueous solution or asa dried form to be used in conjunction with a suitable vehicle such assterile water.

Formulations comprising the aminated psilocybin derivatives of thepresent disclosure for parenteral administration may be formulated to beimmediate and/or modified release. Modified release formulations includedelayed-, sustained-, pulsed-, controlled-, targeted and programmedrelease. Thus the chemical compounds of the disclosure may be formulatedas a solid, semi-solid, or thixotropic liquid for administration as animplanted depot providing modified release of the active compound.Examples of such formulations include drug-coated stents andpoly(dl-lactic-coglycolic) acid (PGLA) microspheres.

The pharmaceutical or recreational drug formulations of the presentdisclosure also may be administered topically to the skin or mucosa,i.e., dermally or transdermally. Example pharmaceutical and recreationaldrug formulations for this purpose include gels, hydrogels, lotions,solutions, creams, ointments, dusting powders, cosmetics, oils, eyedrops, dressings, foams, films, skin patches, wafers, implants, sponges,fibres, bandages and microemulsions. Liposomes may also be used. Examplecarriers include alcohol, water, mineral oil, liquid petrolatum, whitepetrolatum, glycerin, polyethylene glycol and propylene glycol.Penetration enhancers may be incorporate (see: for example, Finnin, B.and Morgan, T. M., 1999 J. Pharm. Sci, 88 (10), 955-958).

Other means of topical administration include delivery byelectroporation, iontophoresis, phonophoresis, sonophoresis andmicroneedle or needle-free (e.g., Powderject™, Bioject™, etc.)injection.

Pharmaceutical and recreational drug formulations for inhalation orinsufflation include solutions and suspensions in pharmaceuticallyacceptable, aqueous, or organic solvents, or mixtures thereof, andpowders. The liquid or solid pharmaceutical compositions can containsuitable pharmaceutically acceptable excipients. In some embodiments,the pharmaceutical compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Pharmaceuticalcompositions in pharmaceutically acceptable solvents can be nebulized byuse of inert gases. Nebulized solutions can be inhaled directly from thenebulizing device or the nebulizing device can be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder pharmaceutical compositions can beadministered, e.g., orally, or nasally, from devices that deliver theformulation in an appropriate manner.

In further embodiments, in which the aminated psilocybin compounds ofpresent disclosure are used as a recreational drug, the compounds may beincluded in compositions such as a food or food product, a beverage, afood seasoning, a personal care product, such as a cosmetic, perfume orbath oil, or oils (both for topical administration as massage oil, or tobe burned or aerosolized). The chemical compounds of the presentdisclosure may also be included in a “vape” product, which may alsoinclude other drugs, such as nicotine, and flavorings.

The pharmaceutical formulations comprising the chemical compounds of thepresent disclosure may be used to treat a subject, and in particular totreat a psychiatric disorder in a subject. Accordingly, the presentdisclosure includes in a further embodiment, a method for treating apsychiatric disorder, the method comprising administering to a subjectin need thereof a pharmaceutical formulation comprising a chemicalcompound or salt thereof having formula (I):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group orN-substituted amino group, and wherein each non-aminated R₂, R₅, R₆, orR₇ is a hydrogen atom, an alkyl group or O-alkyl group, wherein R₄ whenit is not aminated is a hydrogen atom, an alkyl group or O-alkyl group,a hydroxy group, or a phosphate group, and wherein R_(3A) and R_(3B) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group, together with a diluent, carrier, or excipient.

Psychiatric disorders that may be treated include, for example,neurodevelopmental disorders such as intellectual disability, globaldevelopment delay, communication disorders, autism spectrum disorder,and attention-deficit hyperactivity disorder (ADHD); bipolar and relateddisorders, such as mania, and depressive episodes; anxiety disorder,such as generalized anxiety disorder (GAD), agoraphobia, social anxietydisorder, specific phobias (natural events, medical, animal,situational, for example), panic disorder, and separation anxietydisorder; stress disorders, such as acute stress disorder, adjustmentdisorders, post-traumatic stress disorder (PTSD), and reactiveattachment disorder; dissociative disorders, such as dissociativeamnesia, dissociative identity disorder, anddepersonalization/derealization disorder; somatoform disorders, such assomatic symptom disorders, illness anxiety disorder, conversiondisorder, and factitious disorder; eating disorders, such as anorexianervosa, bulimia nervosa, rumination disorder, pica, and binge-eatingdisorder; sleep disorders, such as narcolepsy, insomnia disorder,hypersomnolence, breathing-related sleep disorders, parasomnias, andrestless legs syndrome; disruptive disorders, such as kleptomania,pyromania, intermittent explosive disorder, conduct disorder, andoppositional defiant disorder; depressive disorders, such as disruptivemood dysregulation disorder, major depressive disorder, persistentdepressive disorder (dysthymia), premenstrual dysphoric disorder,substance/medication-induced depressive disorder, postpartum depression,and depressive disorder caused by another medical condition for example,psychiatric and existential distress within life-threatening cancersituations (ACS Pharmacol Transl Sci 4: 553-562; J Psychiatr Res 137:273); substance-related disorders, such as alcohol-related disorders,cannabis related disorders, inhalant-use related disorders, stimulantuse disorders, and tobacco use disorders; neurocognitive disorders, suchas delirium; schizophrenia; compulsive disorders, such as obsessivecompulsive disorders (OCD), body dysmorphic disorder, hoarding disorder,trichotillomania disorder, excoriation disorder, substance/medicationinduced obsessive-compulsive disorder, and obsessive-compulsive disorderrelated to another medical condition; and personality disorders, such asantisocial personality disorder, avoidant personality disorder,borderline personality disorder, dependent personality disorder,histrionic personality disorder, narcissistic personality disorder,obsessive-compulsive personality disorder, paranoid personalitydisorder, schizoid personality disorder, and schizotypal personalitydisorder.

In an aspect, the compounds of the present disclosure may be used to becontacted with a 5-HT_(2A) receptor to thereby modulate the 5-HT_(2A)receptor. Such contacting includes bringing a compound of the presentdisclosure and 5-HT_(2A) receptor together under in vitro conditions,for example, by introducing the compounds in a sample containing a5-HT_(2A) receptor, for example, a sample containing purified 5-HT_(2A)receptors, or a sample containing cells comprising 5-HT_(2A) receptors.In vitro conditions further include the conditions described in Example4 hereof. Contacting further includes bringing a compound of the presentdisclosure and 5-HT_(2A) receptor together under in vivo conditions.Such in vivo conditions include the administration to an animal or humansubject, for example, of a pharmaceutically effective amount of thecompound of the present disclosure, when the compound is formulatedtogether with a pharmaceutically active carrier, diluent, or excipient,as hereinbefore described, to thereby treat the subject. Upon havingcontacted the 5-HT_(2A) receptor, the compound may activate the5-HT_(2A) receptor or inhibit the 5-HT_(2A) receptor.

Thus, in a further aspect, the condition that may be treated inaccordance herewith can be any 5-HT_(2A) receptor mediated disorder.Such disorders include, but are not limited to schizophrenia, psychoticdisorder, attention deficit hyperactivity disorder, autism, and bipolardisorder.

In an aspect, the compounds of the present disclosure may be used to becontacted with a 5-HT_(1A) receptor to thereby modulate the 5-HT_(1A)receptor. Such contacting includes bringing a compound of the presentdisclosure and 5-HT_(1A) receptor together under in vitro conditions,for example, by introducing the compounds in a sample containing a5-HT_(1A) receptor, for example, a sample containing purified 5-HT_(1A)receptors, or a sample containing cells comprising 5-HT_(1A) receptors.In vitro conditions further include the conditions described in Example1 hereof. Contacting further includes bringing a compound of the presentdisclosure and 5-HT_(1A) receptor together under in vivo conditions.Such in vivo conditions include the administration to an animal or humansubject, for example, of a pharmaceutically effective amount of thecompound of the present disclosure, when the compound is formulatedtogether with a pharmaceutically active carrier, diluent, or excipient,as hereinbefore described, to thereby treat the subject. Upon havingcontacted the 5-HT_(2A) receptor, the compound may activate the5-HT_(1A) receptor or inhibit the 5-HT_(1A) receptor.

Thus, in a further aspect, the condition that may be treated inaccordance herewith can be any 5-HT_(1A) receptor mediated disorder.Such disorders include, but are not limited to schizophrenia, psychoticdisorder, attention deficit hyperactivity disorder, autism, and bipolardisorder.

The chemical compounds of the present disclosure may also be used as afeedstock material for other psilocybin derivatives. Thus in oneembodiment, the chemical compounds of the present disclosure may be inused manufacture of a pharmaceutical or recreational drug formulation,wherein the manufacture may comprise derivatizing a chemical compoundhaving the formula

wherein at least one of R₂, R₄, R₅, R₆ or R₇ is an amino group orN-substituted amino group, wherein each non-aminated R₂, R₅, R₆, or R₇is a hydrogen atom, alkyl group or O-alkyl group, wherein R₄ when it isnot aminated is a phosphate group, a hydrogen atom, a hydroxy group, analkyl group, or O-alkyl group, and wherein R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, an aryl group, or an acylgroup, or a salt of the chemical compound.

In order to use the compound having formula (I) as a feedstock, one ormore amino group or N-substituted amino groups may be substituted by anyatoms or groups, for example hydrocarbon groups. Those of skill in theart will be generally familiar with methods that may be used tosubstitute amino group or N-substituted amino groups. In this respect,guidance may be found in Schnepel C. et al. (2017) Chem. Eur. J.23:12064-12086; Durak L. J. et al. (2016) ACS Catal. 6: 1451; RunguphanW. et al. (2013) Org Lett 15: 2850; Corr M. J. et al. (2017) Chem. Sci.8: 2039; and Roy A. D. et al. Chem. Comm. 4831.

Turning now to methods of making the aminated psilocybin derivatives ofthe present disclosure, it is initially noted that the aminatedpsilocybin derivatives of the present disclosure may be prepared in anysuitable manner, including by any organic chemical synthesis methods,biosynthetic methods, or a combination thereof.

One suitable method of making the aminated psilocybin derivatives of thepresent disclosure initially involves selecting and obtaining orpreparing a reactant psilocybin derivative compound, and reacting thecompound under suitable conditions to form an aminated psilocybinderivative.

Suitable reactant psilocybin derivative compounds include compoundscomprising an indole prototype structure (see: FIG. 2 ), including, forexample, a chemical compound having formula (II):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is a reactive groupselected from a nitro group, an azido group, or a hydrogen atom, andwherein R₂, R₄, R₅, R₆, or R₇ which are not a reactive group, are ahydrogen atom, an alkyl of O-alkyl group, and wherein R_(3A) and R_(3B)are each independently a hydrogen atom, an alkyl group, and acyl group,or an aryl group. Reactant psilocybin derivative compound (II) comprisesa plurality of compounds, some examples of which will next be described.

In one example embodiment, the reactant psilocybin derivative can beselected to be a chemical compound wherein R₄ is an O-alkyl group, R₂,R₅, R₆, and R₇ are a hydrogen atom, and R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, an acyl group, or an arylgroup, such as, for example, the reactant psilocybin derivative shown inFIGS. 8A and 8B.

In one example embodiment, the reactant psilocybin derivative can beselected to be a chemical compound wherein R₄ is an O-acyl group, R₂,R₅, R₆, and R₇ are a hydrogen atom, and R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, an acyl group, or an arylgroup, such as, for example, the reactant psilocybin derivative shown inFIGS. 8C and 8D.

In one example embodiment, the reactant psilocybin derivative can beselected to be a chemical compound wherein R₄ is a hydroxyl group, R₂,R₅, R₆, and R₇ are a hydrogen atom, and R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, an acyl group, or an arylgroup, such as, for example, the reactant psilocybin derivative shown inFIG. 8E.

In one example embodiment, the reactant psilocybin derivative can beselected to be a chemical compound wherein R₄ is a phosphate group, R₂,R₅, R₆, and R₇ are a hydrogen atom, and R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, an acyl group, or an arylgroup, such as, for example, the reactant psilocybin derivative shown inFIG. 8F.

In one example embodiment, the reactant psilocybin derivative can beselected to be a chemical compound wherein R₄ is a hydrogen atom, R₂,R₅, R₆, and R₇ are a hydrogen atom, and R_(3A) and R_(3B) are eachindependently a hydrogen atom, an alkyl group, an acyl group, or an arylgroup, such as, for example, the reactant psilocybin derivative shown inFIG. 8G.

The reactant psilocybin derivative compounds may be provided in a moreor less chemically pure form, for example, in the form of a psilocybinderivative preparation having a purity of at least about 95%, at leastabout 96%, at least about 97%, at least about 98%, at least about 99%,or at least 99.9%. The psilocybin derivative may be chemicallysynthesized, or obtained from a fine chemical manufacturer.

In one example embodiment, the reactant psilocybin derivative compound,may be reacted with a nitrogenous compound with a nitrogenous compound,including, for example, a nitrogenous compound selected from nitric acid(HNO₃), a nitrate salt, an acyl nitrate, trifluoromethansulfonylnitrate, nitrosonium tetrafluoroborate (NO₂BF₄), and trifluoracetylnitrate to initially form a nitrated psilocybin compound which can thenbe reacted under reducing conditions to form an aminated psilocybincompound.

Alternatively, in order to form the aminated psilocybin derivatives ofthe present disclosure a nitrated or azido group containing compoundreactant psilocybin compounds may be obtained and reacted under reducingconditions to form the aminated psilocybin derivatives of the presentdisclosure.

Referring now to FIGS. 9A-9B, shown therein is an example reactionwherein 5-nitro-4-O-methyl-psilocybin derivative is converted to the5-amino-4-O-methyl-psilocybin and some example N-substitutedderivatives.

Referring now to FIG. 9A, shown therein is an example chemical reactionwherein nitrosonium tetrafluoroborate (NO₂BF₄) is reacted with a4-O-methyl-psilocybin derivative (FIG. 8A) in a chemical reaction whichresults initially in the formation of an intermediate psilocybinderivative, notably a 4-O-methyl-5-nitro-psilocybin derivative.Subsequently, in the presence of hydrogen, the nitro group of theintermediate psilocybin derivative is reduced to an amino group underthe catalytic hydrogenolysis condition with the help of palladium oncharcoal. This affords the 5-amino-4-O-methyl-psilocybin derivative.

Subsequent N-substitutions on the formed amino group (FIG. 9B) can becarried out using (1) N-acylations, such as N-acetylation with aceticanhydride or N-sulfonylation with sulfur trioxide-pyridine complex, (2)selective N-alkylation via a reaction with an aldehyde followed by areduction of the intermediate imine, such as the reaction withacetaldehyde to form an intermediate imine, followed by a reduction withsodium borohydride.

Referring now to FIG. 9D, shown therein is an example of multistepsynthesis of two 4-O-methyl-psilocybin derivative respectively aminatedat C₅ (compound 9D-8, corresponding with the compound having chemicalformula (IX), set forth herein) and C₇ (compound 9D-10, correspondingwith the compound having chemical formula (XII), set forth herein) using4-methoxyindole (compound 9D-1) as a starting compound. Thus, startingfrom 9D-1, a regioselective 2-nitrovinylation can be carried out using1-(dimethylamino)-2-nitroethylene as an electrophile in the presence oftrifluoroacetic acid as a catalyst. The reaction can provide the desired(E)-3-(2-nitroethenyl)-4-methoxyindole (9D-2) which can be directlyreduced using sodium borohydride in a mixture of ethanol and THF, toprovide the desired 3-(2-nitroethyl)-4-methoxyindole (9D-3), forexample, yielding 33% (2 steps). The nitro group of the side chain canbe further reduced using lithium aluminum hydride in THE to afford theintermediate 4-O-methy-tryptamine (9D-4), for example, yielding 57%. Tofacilitate subsequent nitration, compound 9D-4 can be protected with anexcess of di-tert-butyl decarbonate in the presence of4-N,N-dimethylaminopyridine to provide the tri-Boc-protected4-O-methy-tryptamine (9D-5), for example, yielding 51%. Compound 9D-5can then be subjected to a reaction with benzoyl nitrate, generated bymixing benzoyl chloride and silver nitrate in anhydrous acetonitrile.This can provide three nitrated products respectively at C₂ (compound9D-6a, e.g., 7% yield), C₅ (compound 9D-6b, 14% yield), and C₇ (compound9D-6c, 12%). Compound 9D-6b can subsequently be subjected to a reductionusing ammonium formate in the presence of 10% palladium on charcoal inmethanol at room temperature to afford the 5-aminated intermediate 9D-7(e.g., 88% yield) which can be fully deprotected subsequently by atreatment with trifluoroacetic acid to furnish the desired5-amino-4-O-methyl-psilocybin derivative (9D-8, e.g., 56% yield,corresponding with the compound having chemical formula (IX), set forthherein). Analogously, compound 9D-6c can also subjected to a reductionusing ammonium formate in the presence of 10% palladium on charcoal inmethanol at room temperature to afford the 7-aminated intermediate 9D-9(e.g., 67% yield) which can further be fully deprotected by a treatmentwith trifluoroacetic acid to furnish the desired7-amino-4-O-methyl-psilocybin derivative 9D-10, (e.g., 70% yield,corresponding with the compound having chemical formula (XII), set forthherein).

Thus, referring to the reactant psilocybin derivative compound havingformula (II), the conditions can comprise: (i) appropriately protectingthe side-chain amino group with one or two protecting groups (R_(3a),R_(3b)) along with or without the protection of N₁ using R₁. It is notedthat the protecting groups R₁, R_(3a), R_(3b) can be, for example, analkyl or an acyl group, such as an acetyl group or substituted acetylgroup, such as trifluoroacetyl, or other groups, such as a carbamategroup, e.g., fluorenylmethyloxycarbonyl (Fmoc), benzyloxycarbonyl, ortert-butyloxycarbonyl (Boc) which can, for example, be prepared byreacting with di-tert-butyl dicarbonate in the presence of4-N,N-dimethylaminopyridine (DMAP). It is noted that the protection ofN₁ with R₁ is optional depending on the nature of electrophile which isused in the nitration reaction (referring to FIG. 9D, see: e.g.,reaction 9D-5 to 9D-6a, 9D-6b, 9D-6c), as well as the specific reactionconditions, e.g., pH, temperatures, solvents, catalysts; (ii) reactingthe protected reactant psilocybin compound with a nitrogenous compoundselected from nitric acid (HNO₃); a nitrate salt, such as AgNO₃; an acylnitrate such as trifluoromethansulfonyl nitrate; benzoyl nitrate,nitrosonium tetrafluoroborate (NO₂BF₄), and trifluoracetyl nitrate toform a nitrated compound having chemical formula (XXXI):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is a nitro group, andwherein each non-nitrated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkylgroup or O-alkyl group, wherein R₄ when it is not nitrated is a hydrogenatom, an alkyl group, O-alkyl group, a hydroxy group, or a phosphategroup, and wherein R_(3A) and R_(3B) are a protective group, and whereinR₁ is a protective group or a hydrogen atom, and then (iii) reacting thenitrated compound under a reducing condition to form an aminatedcompound having chemical formula (XXXII):

wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group, andwherein each non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkylgroup or O-alkyl group, wherein R₄ when it is not aminated is a hydrogenatom, an alkyl group, O-alkyl group, a hydroxy group, or a phosphategroup, and wherein R_(3A) and R_(3B) are a protective group, and whereinR₁ is a protective group or a hydrogen atom, then (iv) optionallysubstituting the at least one amino group at R₂, R₄, R₅, R₆, or R₇ groupto form a N-substituted derivative, and then (v) removing the protectinggroups (R₁, R_(3a), R_(3b)) at N₁ and the side chain aminofunctionality, to thereby form a compound having chemical formula (I),and then (vi) optionally substituting the amino group of the side chainto form at least one N-substituted group.

Furthermore, referring now to FIG. 9C, shown therein is an alternativemethod for making the psilocybin derivatives of the present disclosure,notably a direct amination method using hydrogen peroxide and ammoniawith the help of Cu/SiO₂ as a catalyst (T. Yu, R. Yang, S. Xia, G. Li,and C. Hu Catal. Sci. Technol., 2014, 4, 3159-3167). The presence of acatalyst is required in view of the fact that the reaction at practicaltemperatures and pressures is thermodynamically unfavorable. Suchcatalysts may include a platinum-containing catalyst, which may be heldat high temperature, or a reducible metal oxide, for example, oxides ofFe, Ni, Co, Sn, Sb, Bi or Cu, as further described in, for example,Canadian Patent 553,988, and U.S. Pat. Nos. 2,948,755; and 4,031,106.

Similarly, referring further to FIG. 9A, having obtained the nitratedcompound, the reduction reaction is preferably conducted in the presenceof a catalyst capable of providing electrons, for example, a Cu, Sn, Nior Pt containing catalyst. The reduction reaction under acidicconditions may initially lead to the formation of an intermediatepsilocybin derivative possessing a cationic NH₃ ⁺ group, which may bereacted with for example sodium hydroxide to form an amino group or aN-substituted amino group.

Thus, it will be clear that in one embodiment, in an aspect, in thechemical compound having formula (II), at least one of R₂, R₄, R₅, R₆,or R₇ in the reactant psilocybin derivative compound can be a hydrogenatom, and the reaction conditions can comprise reacting the reactantpsilocybin compound with ammonia and hydrogen peroxide in the presenceof a catalyst, such as Cu/SiO₂, to form the chemical compound havingformula (I) and to then optionally substitute the at least one aminogroup in the chemical compound having formula (I) to form at least oneN-substituted group.

Furthermore, referring to FIG. 9D, 1-3 protecting groups can beintroduced to protect the selected substrate compound shown in FIG. 8A,and the protecting groups can be a group different from Boc, such as analkyl and/or acyl group like trifluoroacetyl, trichloroacetyl,dichloroacetyl, chloroacetyl, benzyloxycarbonyl,fluorenylmethyloxycarbonyl (Fmoc) etc. It will now be clear that, in anaspect hereof, the protected reactant psilocybin derivatives disclosedherein may be reacted with a nitrogenous compound, such as, for example,nitric acid (HNO₃), a nitrate salt, an acyl nitrate such astrifluoromethansulfonyl nitrate, benzoyl nitrate and trifluoracetylnitrate, and ammonia to form the aminated psilocybin derivatives of thepresent disclosure. Thus, in addition to reactant psilocybin derivativeshown in FIG. 8A, the example reactant psilocybin derivatives shown inFIGS. 8B-8G may also be reacted with a suitable reagent to form exampleaminated psilocybin derivatives of the present disclosure. The4-O-methyl-psilocybin derivative depicted an FIG. 8A may be reacted toform, for example, the 4-O-methyl-5-amino-psilocybin derivative depictedin FIG. 5A (as already noted), the 4-O-methyl-7-amino-psilocybinderivative depicted in FIG. 6A, and the4-O-methyl-5,7-di-amino-psilocybin depicted in FIG. 7A.

Similarly, the 4-O-ethyl-psilocybin derivative depicted an FIG. 8B maybe reacted in similar reaction sequence to form, for example, the4-O-ethyl-5-amino-psilocybin derivative depicted in FIG. 5B, the4-O-ethyl-7-amino-psilocybin derivative depicted in FIG. 6B, and the4-O-ethyl-5,7-di-amino-psilocybin depicted in FIG. 7B.

Similarly, the 4-acetyl-psilocybin derivative depicted an FIG. 8C may bereacted in similar reaction sequence to form, for example, the4-O-acetyl-5-amino-psilocybin derivative depicted in FIG. 5C, the4-O-acetyl-7-amino-psilocybin derivative depicted in FIG. 6C, and the4-O-acetyl-5,7-di-amino-psilocybin depicted in FIG. 7C.

Similarly, the 4-propanoyl-psilocybin derivative depicted an FIG. 8D maybe reacted in similar reaction sequence to form, for example, the4-O-propanoyl-5-amino-psilocybin derivative depicted in FIG. 5D, the4-O-propanoyl-7-amino-psilocybin derivative depicted in FIG. 6D, and the4-O-propanoyl-5,7-di-amino-psilocybin depicted in FIG. 7D.

Similarly, the 4-hydroxy-psilocybin derivative depicted an FIG. 8E maybe reacted in similar reaction sequence to form, for example, the4-hydroxy-5-amino-psilocybin derivative depicted in FIG. 5E, the4-hydroxy-7-amino-psilocybin derivative depicted in FIG. 6E, and the4-hydroxy-5,7-di-amino-psilocybin depicted in FIG. 7E.

Similarly, the 4-phospho-psilocybin derivative depicted an FIG. 8F maybe reacted in similar reaction sequence to form, for example, the4-phosphate-5-amino-psilocybin derivative depicted in FIG. 5F, the4-phosphate-7-amino-psilocybin derivative depicted in FIG. 6F, and the4-phosphate-5,7-di-amino-psilocybin depicted in FIG. 7F.

Similarly, the psilocybin derivative depicted an FIG. 8G may be reactedto form, for example, the 5-amino-psilocybin derivative depicted in FIG.5G, the 47-amino-psilocybin derivative depicted in FIG. 6G, and the5,7-di-amino-psilocybin depicted in FIG. 7G as well as other analogs.

It is noted that the performance of the reactions, in example differentembodiments, may involve amination of different carbon atoms, i.e., theC₂, C₅, C₆ and/or C₇ atom. In general, reaction conditions may beselected so that different carbon atoms or combinations thereof areaminated. Thus, for example, using either a C₅-nitrated orC₅-azido-substituted psilocybin or derivative as a starting material,the nitro or the azido group can be reduced to afford the5-amino-psilocybin or derivative. The methods can be used to prepare anyother mono-, di- or multi-aminated psilocybin derivatives from theircorresponding nitrated or azido-substituted substrates (see: Kadam, H.K; Tilve, S. G. RSC Adv. 2015, 5, 83391-83407). Typical reductionconditions can be selected from a range of conventional conditions, suchas catalytic hydrogenolysis with the help of heavy metal such aspalladium on charcoal, palladium hydroxide on charcoal, Raney Nickel,platinum oxide; palladium on charcoal with ammonium formate; reactivemetal such as zinc, iron or copper in an acidic media or with an salt,such as zinc/ammonium chloride; organic phosphine such astriphenylphosphine or trimethylphosphine (H.-C, Wu, J.-Q. Yu, J. B.Spencer, Org. Lett., 2004, 6, 4675-4678); sulfur containing reducingagent such as sodium hydrosulfite, sodium sulfide, hydrogen sulfide;tin(I) chloride; organic silanes (R. J. Rahaim, R. E. Maleczka, Jr.,Org. Lett., 2005, 7, 5087-5090). The amination on the psilocybin andderivatives can also be achieved from a precursor substrate containingeither an acyl azide (—CON₃) or amide (—CONH₂) functionality at any ofthe C₂, C₅, C₆, C₇ positions via respectively the Curtis rearrangement(Scriven, E. F. V.; Turnbull, K., Chemical Reviews. 1988, 88, 297-368)or Hoffmann rearrangement (Baumgarten, H.; Smith, H; Stakiis, A, J. Org.Chem 1975, 40 (24): 3554-3561). The obtained amines can be furthersubstituted with N-alkylation or N-acylation or a combination of thetwo, and it can also be modified with a sulfur containing acylatingagent such as sulfur trioxide-pyridine, sulfonyl chloride. Furthermore,the obtained amines can also be reacted with an aldehyde or ketone forform the corresponding imines that can be reduced subsequently.

The reactions may be conducted in any suitable reaction vessel (e.g., atube, bottle). Suitable solvents that may be used are for example,water, alcohol (such as methanol, ethanol, tetrahydrofuran (THF),N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), or a combinationof solvents. Suitable temperatures may range from, for example, e.g.,from about 20° C. to about 100° C. Furthermore, reaction times may bevaried. As will readily be appreciated by those of skill in the art, thereaction conditions may be optimized, for example by preparing severalpsilocybin derivative reactants preparations and azido and reactingthese in different reaction vessels under different reaction conditions,for example, at different temperatures, using different solvents, usingdifferent catalysts etc., evaluating the obtained aminated psilocybinderivative reaction product, adjusting reaction conditions, andselecting a desired reaction condition. Further general guidanceregarding appropriate reaction conditions for performing aminationreactions may be found in, for example Kadarn, H. K.: Tilve, S. G. RSCAdv. 2015, 5, 83391-83407.

In another aspect of the present disclosure, the aminated psilocybincompounds may be made biosynthetically. Accordingly, the presentdisclosure further includes, in one embodiment, a method of making anaminated psilocybin derivative the method comprising:

-   -   (a) contacting a aminated psilocybin precursor compound with a        host cell comprising a psilocybin biosynthetic enzyme        complement, and    -   (b) growing the host cell to produce an aminated psilocybin        derivative or salts thereof having the formula (I):

-   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group        or an N-amino substituted amino group, and wherein each        non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkyl or        O-alkyl group, wherein R₄ when it is not aminated is a hydrogen        atom, an alkyl group or O-alkyl group, a hydroxy group, or a        phosphate group, and wherein R_(3A) and R_(3B) are each        independently a hydrogen atom, an alkyl group, an aryl group, or        an acyl group.

Implementation of the foregoing example embodiment initially involvesproviding aminated psilocybin precursor compounds and host cells havinga psilocybin biosynthetic enzyme complement. Accordingly, next, exampleaminated psilocybin precursor compounds and example host cells that maybe selected and used in accordance with the present disclosure will bedescribed. Thereafter, example methodologies and techniques will bedescribed to contact and use the aminated psilocybin precursor compoundsand cells to produce example aminated psilocybin compounds.

A variety of aminated psilocybin precursor compounds may be selected,prepared, and used. In some embodiments, for example, the aminatedpsilocybin precursor compound is a compound comprising an aminatedindole prototype structure. Examples of such compounds are an aminatedindole, e.g., 2-amino-indole, 4-amino-indole, 5-amino-indole,6-amino-indole, and 7-amino-indole; and aminated tryptophan derivatives,e.g., 2-amino-tryptophan, 4-amino-tryptophan, 5-amino-tryptophan,6-amino-tryptophan, and 7-amino-tryptophan.

Further aminated psilocybin precursor compounds that may be used includeaminated indoles, having the formula (XXIX):

wherein at least one of R₂, R₄, R₅, R₆ and R₇ is an amino group orN-substituted amino group, wherein R₂, R₄, R₅, R₆ and R₇ when they arenot aminated are hydrogen atoms, an alkyl group or O-alkyl group,wherein R₄ when it is not aminated is a hydrogen atom, an alkyl group orO-alkyl group, a hydroxy group, or a phosphate group.

Further aminated psilocybin precursor compounds that may be used includecompounds having the formula (XXVII):

wherein at least one of R₂, R₄, R₅, R₆ and R₇ is an amino group or anN-substituted group, wherein R₂, R₄, R₅, R₆ and R₇ when they are notaminated are hydrogen atoms, an alkyl group or O-alkyl group, wherein R₄when it is not aminated is a hydrogen atom, an alkyl group or O-alkylgroup, a hydroxy group, or a phosphate group

Turning now to the host cells that can be used in accordance with thepresent disclosure, it is initially noted that a variety of host cellsmay be selected in accordance with the present disclosure, includingmicroorganism host cells, plant host cells, and animal host cells.

In accordance herewith the host cell includes a psilocybin biosyntheticenzyme complement. Such cells can be obtained in at least two ways.First, in some embodiments, host cells may be selected in which apsilocybin biosynthetic enzyme complement is naturally present.Generally cells naturally producing psilocybin for example, cells offungal species belonging to the genus psilocybe, are suitable in thisrespect. Second, in some embodiments, a host cell that not naturallyproduces psilocybin may be modulated to produce a psilocybinbiosynthetic enzyme complement. Thus, for example, a nucleic acidsequence encoding a psilocybin biosynthetic enzyme complement may beintroduced into a host cell, and upon cell growth the host cells canmake the psilocybin biosynthetic enzyme complement.

Typically a nucleic acid sequence encoding one or more enzymesconstituting a psilocybin biosynthetic enzyme complement furtherincludes one or more additional nucleic acid sequences, for example, anucleic acid sequences controlling expression of the one or moreenzymes, and these one or more additional nucleic acid sequencestogether with the nucleic acid sequence encoding the one or more enzymescan be said to form a chimeric nucleic acid sequence.

A host cell which upon cultivation expresses the chimeric nucleic acidcan be selected and used in accordance with the present disclosure.Suitable host cells in this respect include, for example, microbialcells, such as bacterial cells, yeast cells, for example, and algalcells or plant cells. A variety of techniques and methodologies tomanipulate host cells to introduce nucleic acid sequences in cells andattain expression exists and are well known to the skilled artisan.These methods include, for example, cation based methods, for example,lithium ion or calcium ion based methods, electroporation, biolistics,and glass beads based methods. As will be known to those of skill in theart, depending on the host cell selected, the methodology to introducenucleic acid material in the host cell may vary, and, furthermore,methodologies may be optimized for uptake of nucleic acid material bythe host cell, for example, by comparing uptake of nucleic acid materialusing different conditions. Detailed guidance can be found, for example,in Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold SpringHarbor Laboratory Press, 2012, Fourth Ed. It is noted that the chimericnucleic acid is a non-naturally occurring chimeric nucleic acid sequenceand can be said to be heterologous to the host cell.

In some embodiments, the one or more enzymes constituting a psilocybinenzyme complement can be selected from by a nucleic acid sequenceselected from the nucleic acid sequences consisting of:

-   -   (a) SEQ. ID NO: 4, SEQ. ID NO: 8, SEQ. ID NO: 11 and SEQ. ID NO:        13;    -   (b) a nucleic acid sequence that is substantially identical to        any one of the nucleic acid sequences of (a);    -   I a nucleic acid sequence that is substantially identical to any        one of the nucleic acid sequences of (a) but for the        degeneration of the genetic code;    -   (d) a nucleic acid sequence that is complementary to any one of        the nucleic acid sequences of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 5, SEQ.        ID NO: 9, SEQ. ID NO: 12 and SEQ. ID NO: 14;    -   (f) a nucleic acid sequence that encodes a functional variant of        any one of the amino acid sequences set forth in SEQ. ID NO: 5,        SEQ. ID NO: 9, SEQ. ID NO: 12 and SEQ. ID NO: 14; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f).

Thus any of the nucleic acid sequence set forth in (a), (b), (c), (d),(e), (f) or (g) may be selected and introduced into a host cell. Inparticular, however the nucleic acid sequence is selected in conjunctionwith the selected psilocybin precursor compound, as hereinafter furtherdiscussed in reference with FIG. 10 .

One example host cell that conveniently may be used is Escherichia coli.The preparation of the E. coli vectors may be accomplished usingcommonly known techniques such as restriction digestion, ligation, gelelectrophoresis, DNA sequencing, the polymerase chain reaction (PCR) andother methodologies. A wide variety of cloning vectors is available toperform the necessary steps required to prepare a recombinant expressionvector. Among the vectors with a replication system functional in E.coli, are vectors such as pBR322, the pUC series of vectors, the M13 mpseries of vectors, pBluescript etc. Suitable promoter sequences for usein E. coli include, for example, the T7 promoter, the T5 promoter,tryptophan (trp) promoter, lactose (lac) promoter, tryptophan/lactose(tac) promoter, lipoprotein (lpp) promoter, and A phage PL promoter.Typically, cloning vectors contain a marker, for example, an antibioticresistance marker, such as ampicillin or kanamycin resistance marker,allowing selection of transformed cells. Nucleic acid sequences may beintroduced in these vectors, and the vectors may be introduced in E.coli by preparing competent cells, electroporation or using other wellknown methodologies to a person of skill in the art. E. coli may begrown in an appropriate medium, such as Luria-Broth medium andharvested. Recombinant expression vectors may readily be recovered fromcells upon harvesting and lysing of the cells.

Another example host cell that may be conveniently used is a yeast cell.Example yeast host cells that can be used are yeast cells belonging tothe genus Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces,Pichia, Hansenuia, and Yarrowia. In specific example embodiments, theyeast cell can be a Saccharomyces cerevisiae cell, a Yarrowia lipolyticacell, or Pichia pastoris cell.

A number of vectors exist for the expression of recombinant proteins inyeast host cells. Examples of vectors that may be used in yeast hostcells include, for example, Yip type vectors, YEp type vectors, YRp typevectors, YCp type vectors, pGPD-2, pAO815, pGAPZ, pGAPZα, pHIL-D2,pHIL-S1, pPIC3.5K, pPIC9K, pPICZ, pPICZα, pPIC3K, pHWO10, pPUZZLE and 2μm plasmids. Such vectors are known to the art and are, for example,described in Cregg et al, Mol Biotechnol. (2000) 16(1): 23-52. Suitablepromoter sequences for use in yeast host cells are also known anddescribed, for example, in Mattanovich et al., Methods Mol Biol, 2012,824:329-58, and in Romanos et al, 1992, Yeast 8: 423-488. Examples ofsuitable promoters for use in yeast host cells include promoters ofglycolytic enzymes, like triosephosphate isomerase (TPI),phosphoglycerate kinase (PGK), glyceraidehyde-3-phosphate dehydrogenase(GAPDH or GAP) and variants thereof, lactase (LAC) and galactosidase(GAL), P. pastoris glucose-6-phosphate isomerase promoter (PPGI), the3-phosphoglycerate kinase promoter (PPGK), the glycerol aldehydephosphate dehydrogenase promoter (PGAP), translation elongation factorpromoter (PTEF), S. cerevisiae enolase (ENO-1), S. cerevisiaegalactokinase (GAL1), S. cerevisiae alcoholdehydrogenase/glyceraldehyde-3-phosphate dehydrogenase (ADH1, ADH2/GAP),S. cerevisiae triose phosphate isomerase (TPI), S. cerevisiaemetallothionein (CUP1), and S. cerevisiae 3-phosphoglycerate kinase(PGK), and the maltase gene promoter (MAL). Marker genes suitable foruse in yeast host cells are also known to the art. Thus, antibioticresistance markers, such as ampicillin resistance markers, can be usedin yeast, as well as marker genes providing genetic functions foressential nutrients, for example, leucine (LEU2), tryptophan (TRP1 andTRP2), uracil (URA3, URA5, URA6), histidine (HIS3), and the like.Methods for introducing vectors into yeast host cells can, for example,be found in S. Kawai et al., 2010, Bioeng. Bugs 1(6): 395-403.

Further, guidance with respect to the preparation of expression vectorsand introduction thereof into host cells, including in E. coli cells,yeast cells, and other host cells, may be found in, for example:Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold SpringHarbor Laboratory Press, 2012, Fourth Ed.

Thus, to briefly recap, a host cell comprising a chimeric nucleic acidcomprising (i) a nucleic acid sequence controlling expression in a hostcell and (ii) a nucleic acid sequence encoding a psilocybin biosyntheticenzyme complement, can be prepared in accordance with the presentdisclosure.

In accordance herewith, host cells are grown to multiply and to expressa chimeric nucleic acid. Expression of the chimeric nucleic acid resultsin the biosynthetic production in the host cell of a psilocybinbiosynthetic enzyme complement. Growth media and growth conditions canvary depending on the host cell that is selected, as will be readilyappreciated to those of ordinary skill in the art. Growth mediatypically contain a carbon source, one or several nitrogen sources,essential salts including salts of potassium, sodium, magnesium,phosphate and sulphate, trace metals, water soluble vitamins, andprocess aids including but not limited to antifoam agents, proteaseinhibitors, stabilizers, ligands, and inducers. Example carbon sourcesare e.g., mono- or disaccharides. Example nitrogen sources are, e.g.,ammonia, urea, amino acids, yeast extract, corn steep liquor and fullyor partially hydrolyzed proteins. Example trace metals are e.g., Fe, Zn,Mn, Cu, Mo and H₃BOs. Example water soluble vitamins are e.g. biotin,pantothenate, niacin, thiamine, p-aminobenzoic acid, choline,pyridoxine, folic acid, riboflavin, and ascorbic acid. Further, specificexample media include liquid culture media for the growth of yeast cellsand bacterial cells including, Luria-Bertani (LB) broth for bacterialcell cultivation, and yeast extract peptone dextrose (YEPD or YPD), foryeast cell cultivation. Further media and growth conditions can be foundin Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold SpringHarbor Laboratory Press, 2012, Fourth Ed.

In order for the host cells to produce the aminated psilocybin compound,the cells are provided with a precursor compound. Thus in accordanceherewith, host cells may be contacted with a psilocybin precursorcompound. In some embodiments, a psilocybin precursor compound can beexogenously supplied, for example, by including a psilocybin precursorcompound in the growth medium of the host cells, and growing the hostcells in a medium including the psilocybin precursor compound.

Referring next to FIG. 10 , shown therein is an example biosyntheticpathway showing the conversion of example psilocybin precursor compoundsto form an aminated psilocybin. Thus, as can be appreciated from FIG. 10, various psilocybin precursor compounds may be selected and prepared inaminated form, in conjunction with a psilocybin biosynthetic enzymecomplement. Thus, by way of example, aminated tryptophan (e.g., 2-,5-,6-, or 7-aminated tryptophan) may be selected and contacted with a hostcell comprising a psilocybin biosynthetic enzyme complement comprisingtryptophan decarboxylase and optionally N-acetyl transferase, and upongrowth of the cells aminated psilocybin derivatives can be formed. Byway of further example, aminated indole (e.g., 2-,5-, 6-, or 7-aminatedindole) may be selected and contacted with a host cell comprising apsilocybin biosynthetic enzyme complement comprising tryptophan synthasesubunit B polypeptide and tryptophan decarboxylase and optionallyN-acetyl transferase, and upon growth of the cells aminated psilocybinderivatives can be formed

In some embodiments, the psilocybin precursor compound can be anaminated psilocybin precursor compound which is exogenously supplied toa host cell, for example by inclusion in the host cell's growth medium.Thus, for example, referring to FIG. 10 , it will be understood that inaccordance herewith, for example, 7-amino-indole or 7-amino-tryptophan,may be included in the growth medium of a host cell comprising apsilocybin biosynthetic enzyme complement.

Referring to FIG. 10 , in a further example embodiment, the aminatedpsilocybin precursor compound can be an aminated indole, having theformula (XXIX):

-   -   wherein at least one of R₂, R₄, R₅, R₆ and R₇ is an amino group        or N-substituted amino group, wherein R₂, R₄, R₅, R₆ and R₇ when        they are not aminated are hydrogen atoms, an alkyl group or        O-alkyl group, wherein R₄ when it is not aminated is a hydrogen        atom, an alkyl, O-alkyl or O-aryl group, a hydroxy group, or a        phosphate group,        the psilocybin biosynthetic enzyme complement can comprise:    -   (i) a tryptophan synthase subunit B polypeptide encoded by a        nucleic acid selected from:        -   (a) SEQ. ID NO: 8;        -   (b) a nucleic acid sequence that is substantially identical            to the nucleic acid sequence of (a);        -   (c) a nucleic acid sequence that is substantially identical            to the nucleic acid sequence of (a) but for the degeneration            of the genetic code;        -   (d) a nucleic acid sequence that is complementary to the            nucleic acid sequence of (a);        -   (e) a nucleic acid sequence encoding a polypeptide having            any one of the amino acid sequences set forth in SEQ. ID NO:            9;        -   (f) a nucleic acid sequence that encodes a functional            variant of any one of the amino acid sequences set forth in            SEQ. ID NO: 9; and        -   (g) a nucleic acid sequence that hybridizes under stringent            conditions to any one of the nucleic acid sequences set            forth in (a), (b), (c), (d), (e) or (f); and    -   (ii) a tryptophan decarboxylase encoded by a nucleic acid        sequence selected from:        -   (a) SEQ. ID NO: 11;        -   (b) a nucleic acid sequence that is substantially identical            to the nucleic acid sequence of (a);        -   (c) a nucleic acid sequence that is substantially identical            to the nucleic acid sequence of (a) but for the degeneration            of the genetic code;        -   (d) a nucleic acid sequence that is complementary to the            nucleic acid sequence of (a);        -   (e) a nucleic acid sequence encoding a polypeptide having            any one of the amino acid sequences set forth in SEQ. ID NO:            12;        -   (f) a nucleic acid sequence that encodes a functional            variant of any one of the amino acid sequences set forth in            SEQ. ID NO: 12; and        -   (g) a nucleic acid sequence that hybridizes under stringent            conditions to any one of the nucleic acid sequences set            forth in (a), (b), (c), (d), (e) or (f); and the formed            aminated psilocybin derivative can be a compound having            formula (XXVIII):

-   -   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino            group or N-substituted amino group, and wherein each            non-aminated R₂, R₅, R₆, or R₇ is a hydrogen atom, an alkyl            group or O-alkyl group, wherein R₄ when it is not aminated            is a hydrogen atom, an alkyl group or O-alkyl group, a            hydroxy group, or a phosphate group, and wherein at least            one of R_(3A) and R_(3B) are hydrogen atom.

Referring further to FIG. 10 , in another example embodiment, theaminated psilocybin precursor compound can be a compound, having theformula (XXVII):

-   -   wherein at least one of R₂, R₄, R₅, R₆ and R₇ is an amino group        or an N-substituted amino group, wherein R₂, R₄, R₅, R₆ and R₇        when they are not aminated are hydrogen atoms, an alkyl group or        O-alkyl group, wherein R₄ when it is not aminated is a hydrogen        atom, an alkyl group or O-alkyl group, a hydroxy group, or a        phosphate group;        the psilocybin biosynthetic enzyme complement can comprise:        a tryptophan decarboxylase encoded by a nucleic acid sequence        selected from:    -   (a) SEQ. ID NO: 11;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 12;    -   (f) a nucleic acid sequence that encodes a functional variant of        any one of the amino acid sequences set forth in SEQ. ID NO: 12;        and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f); and the formed aminated        psilocybin derivative can be a compound having formula (XXVIII):

-   -   wherein at least one of R₂, R₄, R₅, R₆, or R₇ is an amino group        or N-substituted amino group, and wherein each non-aminated R₂,        R₅, R₆, or R₇ is a hydrogen atom, an alkyl group or O-alkyl        group, wherein R₄ when it is not aminated is a hydrogen atom, an        alkyl group or O-alkyl group, a hydroxy group, or a phosphate        group, and wherein at least one of R_(3A) and R_(3B) are        hydrogen atom.

In some embodiments, in formula (XXVIII) R_(3A) and R_(3B) are each ahydrogen atom.

Referring again to FIG. 10 , the psilocybin biosynthetic enzymecomplement can, in addition to the aforementioned tryptophandecarboxylase and tryptophan synthase subunit B polypeptide furthercomprise an N-acetyl transferase.

In at least one embodiment, in an aspect, the N-acetyl transferase canbe an enzyme encoded by. a nucleic acid sequence selected from:

-   -   (a) SEQ. ID NO: 4;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 5;    -   (f) a nucleic acid sequence that encodes a functional variant of        any one of the amino acid sequences set forth in SEQ. ID NO: 5;        and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f).

In at least one embodiment, in an aspect, the formed aminated psilocybincompound can have the formula (XXX):

-   -   wherein at least one of R₂, R₄, R₅, R₆ or R₇ is an amino group        or N-substituted amino group, wherein each non-aminated R₂, R₅,        R₆, or R₇ is a hydrogen atom, an alkyl group or O-alkyl group,        wherein R₄ when it is not aminated is a phosphate group, a        hydrogen atom or an alkyl group or O-alkyl group.

Referring again to FIG. 10 , the psilocybin biosynthetic enzymecomplement can, in addition to the aforementioned tryptophandecarboxylase and tryptophan synthase subunit B polypeptide furthercomprise an N-methyl transferase.

In at least one embodiment, in an aspect, the N-methyl transferase canbe an enzyme encoded by a nucleic acid sequence selected from:

-   -   (a) SEQ. ID NO: 13;    -   (b) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a);    -   (c) a nucleic acid sequence that is substantially identical to        the nucleic acid sequence of (a) but for the degeneration of the        genetic code;    -   (d) a nucleic acid sequence that is complementary to the nucleic        acid sequence of (a);    -   (e) a nucleic acid sequence encoding a polypeptide having any        one of the amino acid sequences set forth in SEQ. ID NO: 14;    -   (f) a nucleic acid sequence that encodes a functional variant of        the amino acid sequence set forth in SEQ. ID NO: 14; and    -   (g) a nucleic acid sequence that hybridizes under stringent        conditions to any one of the nucleic acid sequences set forth in        (a), (b), (c), (d), (e) or (f).

In at least one embodiment, in an aspect, the formed aminated psilocybincompound can have the chemical formula (XXXIII):

-   -   wherein at least one of R₂, R₄, R₅, R₆ or R₇ is an amino group        or substituted amino group, wherein each non-aminated R₂, R₅,        R₆, or R₇ is a hydrogen atom, or an alkyl group or O-alkyl        group, wherein R₄ when it is not aminated is a phosphate group,        a hydrogen atom or an alkyl group or O-alkyl group, and wherein        at least one of R_(3a) and R_(3b) is an amino group, and wherein        a non-aminated R_(3a) and R_(3b) is a hydrogen atom.

It will be clear to those of skill in the art that a significant varietyof different aminated psilocybin precursor compounds may be selected.FIG. 10 in this respect provides guidance and allows a person of skillin the art to select appropriate psilocybin precursor compounds and amatching a psilocybin biosynthetic enzyme complement.

Upon production by the host cells of the aminated psilocybin compoundsin accordance with the methods of the present disclosure, the aminatedpsilocybin compounds may be extracted from the host cell suspension, andseparated from other constituents within the host cell suspension, suchas media constituents and cellular debris. Separation techniques will beknown to those of skill in the art and include, for example, solventextraction (e.g., butane, chloroform, ethanol), column chromatographybased techniques, high-performance liquid chromatography (HPLC), forexample, and/or countercurrent separation (CCS) based systems. Therecovered aminated psilocybin compounds may be obtained in a more orless pure form, for example, a preparation of aminated psilocybincompounds of at least about 60% (w/w), about 70% (w/w), about 80% (w/w),about 90% (w/w), about 95% (w/w), about 96% (w/w), about 97% (w/w),about 98% (w/w) or about 99% (w/w) purity may be obtained. Thus, in thismanner, aminated psilocybin derivatives in more or less pure form may beprepared.

Similarly, other methods of making the aminated psilocybin compoundsthat may be used in accordance herewith may yield preparations ofaminated compounds of at least about 60% (w/w), about 70% (w/w), about80% (w/w), about 90% (w/w), about 95% (w/w), about 96% (w/w), about 97%(w/w), about 98% (w/w), or about 99% (w/w) purity.

It will now be clear form the foregoing that novel aminated psilocybinderivatives are disclosed herein. The aminated psilocybin compounds maybe formulated for use as a pharmaceutical drug or recreational drug. Theaminated psilocybin compounds may also be used as a feedstock to produceother psilocybin derivatives.

Hereinafter are provided examples of specific implementations forperforming the methods of the present disclosure, as well asimplementations representing the compositions of the present disclosure.The examples are provided for illustrative purposes only, and are notintended to limit the scope of the present disclosure in any way.

Summary of Sequences

SEQ. ID NO: 1 sets forth a nucleic acid sequence of pCDM4 vector.

SEQ. ID NO: 2 sets forth a nucleic acid sequence encoding a syntheticFLAG epitope tag polypeptide.

SEQ. ID NO: 3 sets forth deduced amino acid sequence of a synthetic FLAGepitope tag polypeptide.

SEQ. ID NO: 4 sets forth a nucleic acid sequence encoding a Streptomycesgriseofuscus PsmF N-acetyltransferase polypeptide.

SEQ. ID NO: 5 sets forth a deduced amino acid sequence of a Streptomycesgriseofuscus PsmF N-acetyltransferase polypeptide.

SEQ. ID NO: 6 sets forth a nucleic acid sequence encoding a synthetic V5epitope tag polypeptide.

SEQ. ID NO: 7 sets forth deduced amino acid sequence of a synthetic V5epitope tag polypeptide.

SEQ. ID NO: 8 sets forth a nucleic acid sequence encoding a mutatedThermotoga maritima TmTrpB-2F3 tryptophan synthase subunit Bpolypeptide.

SEQ. ID NO: 9 sets forth a deduced amino acid sequence of a mutatedThermotoga maritima TmTrpB-2F3 tryptophan synthase subunit Bpolypeptide.

SEQ. ID NO: 10 sets forth a nucleic acid sequence of pETM6-H10 vector

SEQ. ID NO: 11 sets forth a nucleic acid sequence encoding a Bacillusatrophaeus BaTDC tryptophan decarboxylase polypeptide.

SEQ. ID NO: 12 sets forth a deduced amino acid sequence of a Bacillusatrophaeus BaTDC tryptophan decarboxylase polypeptide.

SEQ. ID NO: 13 sets forth an Escherichia coli codon optimized nucleicacid sequence encoding a Rhinella marina N-methyltransferasepolypeptide.

SEQ. ID NO: 14 sets forth a deduced amino acid sequence of a Rhinellamarina N-methyltransferase polypeptide.

SEQUENCES SEQ. ID NO: 1GCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGATGCTCCACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCGCCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGCCTAGGATCGAGATCGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGCAGATCTCAATTGGATATCGGCCGGCCACGCGATCGCTGACGTCGGTACCCTCGAGTCTGGTAAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCGTCTACTAGTCGCAGCTTAATTAACCTAAACTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTAGCGAAAGGAGGAGTCGACACTGCTTCCGGTAGTCAATAAACCGGTAAACCAGCAATAGACATAAGCGGCTATTTAACGACCCTGCCCTGAACCGACGACCGGGTCATCGTGGCCGGATCTTGCGGCCCCTCGGCTTGAACGAATTGTTAGACATTATTTGCCGACTACCTTGGTGATCTCGCCTTTCACGTAGTGGACAAATTCTTCCAACTGATCTGCGCGCGAGGCCAAGCGATCTTCTTCTTGTCCAAGATAAGCCTGTCTAGCTTCAAGTATGACGGGCTGATACTGGGCCGGCAGGCGCTCCATTGCCCAGTCGGCAGCGACATCCTTCGGCGCGATTTTGCCGGTTACTGCGCTGTACCAAATGCGGGACAACGTAAGCACTACATTTCGCTCATCGCCAGCCCAGTCGGGCGGCGAGTTCCATAGCGTTAAGGTTTCATTTAGCGCCTCAAATAGATCCTGTTCAGGAACCGGATCAAAGAGTTCCTCCGCCGCTGGACCTACCAAGGCAACGCTATGTTCTCTTGCTTTTGTCAGCAAGATAGCCAGATCAATGTCGATCGTGGCTGGCTCGAAGATACCTGCAAGAATGTCATTGCGCTGCCATTCTCCAAATTGCAGTTCGCGCTTAGCTGGATAACGCCACGGAATGATGTCGTCGTGCACAACAATGGTGACTTCTACAGCGCGGAGAATCTCGCTCTCTCCAGGGGAAGCCGAAGTTTCCAAAAGGTCGTTGATCAAAGCTCGCCGCGTTGTTTCATCAAGCCTTACGGTCACCGTAACCAGCAAATCAATATCACTGTGTGGCTTCAGGCCGCCATCCACTGCGGAGCCGTACAAATGTACGGCCAGCAACGTCGGTTCGAGATGGCGCTCGATGACGCCAACTACCTCTGATAGTTGAGTCGATACTTCGGCGATCACCGCTTCCCTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGCCAGCTCACTCGGTCGCTACGCTCCGGGCGTGAGACTGCGGCGGGCGCTGCGGACACATACAAAGTTACCCACAGATTCCGTGGATAAGCAGGGGACTAACATGTGAGGCAAAACAGCAGGGCCGCGCCGGTGGCGTTTTTCCATAGGCTCCGCCCTCCTGCCAGAGTTCACATAAACAGACGCTTTTCCGGTGCATCTGTGGGAGCCGTGAGGCTCAACCATGAATCTGACAGTACGGGCGAAACCCGACAGGACTTAAAGATCCCCACCGTTTCCGGCGGGTCGCTCCCTCTTGCGCTCTCCTGTTCCGACCCTGCCGTTTACCGGATACCTGTTCCGCCTTTCTCCCTTACGGGAAGTGTGGCGCTTTCTCATAGCTCACACACTGGTATCTCGGCTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTAAGCAAGAACTCCCCGTTCAGCCCGACTGCTGCGCCTTATCCGGTAACTGTTCACTTGAGTCCAACCCGGAAAAGCACGGTAAAACGCCACTGGCAGCAGCCATTGGTAACTGGGAGTTCGCAGAGGATTTGTTTAGCTAAACACGCGGTTGCTCTTGAAGTGTGCGCCAAAGTCCGGCTACACTGGAAGGACAGATTTGGTTGCTGTGCTCTGCGAAAGCCAGTTACCACGGTTAAGCAGTTCCCCAACTGACTTAACCTTCGATCAAACCACCTCCCCAGGTGGTTTTTTCGTTTACAGGGCAAAAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACTGAACCGCTCTAGATTTCAGTGCAATTTATCTCTTCAAATGTAGCACCTGAAGTCAGCCCCATACGATATAAGTTGTAATTCTCATGTTAGTCATGCCCCGCGCCCACCGGAAGGAGCTGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTCGGTATCGTCGTATCCCACTACCGAGATGTCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGGCAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGAATTTGATTGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGAACTTAATGGGCCC SEQ. ID NO: 2GACTACAAGGATGACGATGACAAA SEQ. ID NO: 3 DYKDDDDK SEQ. ID NO: 4ATGAACACCTTCAGAACAGCCACTGCCAGAGACATACCTGATGTAGCAGCAACTCTTACGGAAGCCTTCGCAACTGATCCACCCACGCAGTGGGTGTTCCCCGACGGTACTGCCGCCGTCAGCAGGTTCTTTACACATGTTGCAGATAGGGTTCACACGGCCGGTGGTATTGTTGAGCTACTACCAGACAGAGCCGCCATGATTGCATTGCCACCACACGTGAGGCTGCCAGGAGAAGCTGCCGACGGAAGGCAGGCGGAAATTCAGAGAAGGCTGGCAGACAGGCACCCGCTGACACCTCACTACTACCTGCTGTTTTACGGAGTTAGAACGGCACACCAGGGTTCGGGATTGGGCGGAAGAATGCTGGCCAGATTAACTAGCAGAGCTGATAGGGACAGGGTGGGTACATATACTGAGGCATCCACCTGGCGTGGCGCTAGACTGATGCTGAGACATGGATTCCATGCTACAAGGCCACTAAGATTGCCAGATGGACCCAGCATGTTTCCACTTTGGAGAGATCCAATCCATGATCATTCTGATTAG SEQ. ID NO: 5MNTFRTATARDIPDVAATLTEAFATDPPTQWVFPDGTAAVSRFFTHVADRVHTAGGIVELLPDRAAMIALPPHVRLPGEAADGRQAEIQRRLADRHPLTPHYYLLFYGVRTAHQGSGLGGRMLARLTSRADRDRVGTYTEASTWRGARLMLRHGFHATRPLRLPDGPSMFPLWRDPI HDHSDSEQ. ID NO: 6 GGTAAGCCAATTCCAAATCCTTTGTTGGGTTTGGACTCCACC SEQ. ID NO: 7GKPIPNPLLGLDST SEQ. ID NO: 8ATGAAAGGATATTTCGGACCATACGGTGGCCAGTACGTACCAGAAATATTAATGGGTGCCTTAGAGGAGTTAGAGGCAGCATACGAGGAGATTATGAAGGATGAGAGCTTCTGGAAGGAGTTCAACGATCTACTGAGGGATTACGCAGGCAGACCAACGCCATTGTACTTTGCCAGGAGATTGTCTGAGAAGTACGGCGCCCGTGTTTACTTGAAGCGTGAGGATCTGCTGCACACTGGAGCACACAAGATAAATAACGCTATCGGACAGGTTTTATTGGCCAAATTAATGGGGAAGACACGTATCATAGCCGAGACGGGAGCTGGGCAGCATGGAGTCGCTACTGCTACCGCTGCTGCCCTGTTCGGAATGGAATGTGTGATCTACATGGGTGAAGAGGACACAATCAGACAGAAGTTGAACGTGGAGCGTATGAAATTATTAGGGGCTAAAGTTGTCCCTGTTAAGTCTGGCAGTAGGACCTTGAAGGATGCGATAGACGAGGCTTTGAGAGACTGGATTACTAATTTACAGACAACATATTATGTTATCGGATCTGTTGTTGGTCCCCACCCTTACCCAATTATCGTAAGGAATTTCCAGAAGGTTATCGGTGAGGAGACCAAGAAGCAAATACCAGAAAAGGAAGGTCGTTTGCCAGACTATATAGTTGCCTGCGTAGGCGGCGGTAGCAATGCCGCAGGTATATTTTACCCATTCATAGACTCTGGAGTAAAGCTGATAGGTGTTGAGGCAGGTGGCGAGGGATTGGAGACAGGTAAACACGCAGCCTCGTTATTAAAGGGTAAAATTGGCTATTTACATGGATCGAAGACCTTTGTTCTACAAGATGACTGGGGTCAAGTCCAAGTGAGCCATTCGGTGTCAGCTGGTCTTGACTATTCAGGAGTAGGACCTGAGCATGCTTATTGGAGAGAGACAGGGAAGGTTCTGTACGACGCAGTGACTGACGAAGAGGCTTTGGACGCATTTATAGAGTTATCAAGACTAGAGGGCATTATACCCGCTTTAGAGTCATCGCATGCTCTAGCATATTTGAAGAAGATAAATATAAAAGGTAAGGTTGTGGTGGTCAACCTATCAGGGAGAGGGGATAAAGACCTGGAGTCAGTCTTAAACCATCCATACGTGAGAGAAAGAATTAGATGA SEQ. ID NO: 9MKGYFGPYGGQYVPEILMGALEELEAAYEEIMKDESFWKEFNDLLRDYAGRPTPLYFARRLSEKYGARVYLKREDLLHTGAHKINNAIGQVLLAKLMGKTRIIAETGAGQHGVATATAAALFGMECVIYMGEEDTIRQKLNVERMKLLGAKVVPVKSGSRTLKDAIDEALRDWITNLQTTYYVIGSVVGPHPYPIIVRNFQKVIGEETKKQIPEKEGRLPDYIVACVGGGSNAAGIFYPFIDSGVKLIGVEAGGEGLETGKHAASLLKGKIGYLHGSKTFVLQDDWGQVQVSHSVSAGLDYSGVGPEHAYWRETGKVLYDAVTDEEALDAFIELSRLEGIIPALESSHALAYLKKINIKGKVVVVNLSGRGDKDLESVLNHPYVRERIR SEQ. ID NO: 10GAAGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGCAGATCTCAATTGGATATCGGCCGGCCACGCGATCGCTGACGTCGGTACCCTCGAGTCTGGTAAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCGTCTACTAGTCGCAGCTTAATTAACCTAAACTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTAGCGAAAGGAGGAGTCGACTATATCCGGATTGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCTGGCGGCACGATGGCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATCATGATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGAAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGGACCAGAGAAAAATCACTCAGGGTCAATGCCAGCGCTTCGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCAGCAGCATCCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGTTTCCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGTCGCAGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGATTCATTCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACGACAGGAGCACGATCATGCTAGTCATGCCCCGCGCCCACCGGAAGGAGCTGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTCGGTATCGTCGTATCCCACTACCGAGATGTCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGGCAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGAATTTGATTGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGAACTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGATGCTCCACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCGCCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGCCTAGGATCGAGATCGATCTCGATCCCGCGAAATTAATAC GACTCACTACGSEQ. ID NO: 11ATGATGTCTGAAAATTTGCAATTGTCAGCTGAAGAAATGAGACAATTGGGTTACCAAGCAGTTGATTTGATCATCGATCACATGAACCATTTGAAGTCTAAGCCAGTTTCAGAAACAATCGATTCTGATATCTTGAGAAATAAGTTGACTGAATCTATCCCAGAAAATGGTTCAGATCCAAAGGAATTGTTGCATTTCTTGAACAGAAACGTTTTTAATCAAATTACACATGTTGATCATCCACATTTCTTGGCTTTTGTTCCAGGTCCAAATAATTACGTTGGTGTTGTTGCAGATTTCTTGGCTTCTGGTTTTAATGTTTTTCCAACTGCATGGATTGCTGGTGCAGGTGCTGAACAAATCGAATTGACTACAATTAATTGGTTGAAATCTATGTTGGGTTTTCCAGATTCAGCTGAAGGTTTATTTGTTTCTGGTGGTTCAATGGCAAATTTGACAGCTTTGACTGTTGCAAGACAGGCTAAGTTGAACAACGATATCGAAAATGCTGTTGTTTACTTCTCTGATCAAACACATTTCTCAGTTGATAGAGCATTGAAGGTTTTAGGTTTTAAACATCATCAAATCTGTAGAATCGAAACAGATGAACATTTGAGAATCTCTGTTTCAGCTTTGAAGAAACAAATTAAAGAAGATAGAACTAAGGGTAAAAAGCCATTCTGTGTTATTGCAAATGCTGGTACTACAAATTGTGGTGCTGTTGATTCTTTGAACGAATTAGCAGATTTGTGTAACGATGAAGATGTTTGGTTGCATGCTGATGGTTCTTATGGTGCTCCAGCTATCTTGTCTGAAAAGGGTTCAGCTATGTTGCAAGGTATTCATAGAGCAGATTCTTTGACTTTAGATCCACATAAGTGGTTGTTCCAACCATACGATGTTGGTTGTGTTTTGATCAGAAACTCTCAATATTTGTCAAAGACTTTTAGAATGATGCCAGAATACATCAAGGATTCAGAAACTAACGTTGAAGGTGAAATTAATTTCGGTGAATGTGGTATCGAATTGTCAAGAAGATTCAGAGCTTTGAAGGTTTGGTTGTCTTTTAAAGTTTTCGGTGTTGCTGCTTTTAGACAAGCAATCGATCATGGTATCATGTTAGCAGAACAAGTTGAAGCATTTTTGGGTAAAGCAAAAGATTGGGAAGTTGTTACACCAGCTCAATTGGGTATCGTTACTTTTAGATACATTCCATCTGAATTGGCATCAACAGATACTATTAATGAAATTAATAAGAAATTGGTTAAGGAAATCACACATAGAGGTTTCGCTATGTTATCTACTACAGAATTGAAGGAAAAGGTTGTTATTAGATTGTGTTCAATTAATCCAAGAACTACAACTGAAGAAATGTTGCAAATCATGATGAAGATTAAAGCATTGGCTGAAGAAGTTTCTATTTCATACCCATGTGTTGCTGAATAA SEQ. ID NO: 12MMSENLQLSAEEMRQLGYQAVDLIIDHMNHLKSKPVSETIDSDILRNKLTESIPENGSDPKELLHFLNRNVFNQITHVDHPHFLAFVPGPNNYVGVVADFLASGFNVFPTAWIAGAGAEQIELTTINWLKSMLGFPDSAEGLFVSGGSMANLTALTVARQAKLNNDIENAVVYFSDQTHFSVDRALKVLGFKHHQICRIETDEHLRISVSALKKQIKEDRTKGKKPFCVIANAGTTNCGAVDSLNELADLCNDEDVWLHADGSYGAPAILSEKGSAMLQGIHRADSLTLDPHKWLFQPYDVGCVLIRNSQYLSKTFRMMPEYIKDSETNVEGEINFGECGIELSRRFRALKVWLSFKVFGVAAFRQAIDHGIMLAEQVEAFLGKAKDWEVVTPAQLGIVTFRYIPSELASTDTINEINKKLVKEITHRGFAMLSTTELKEKVVIRLCSINPRTTTEEMLQIMMKIKALAEEV SISYPCVAESEQ. ID NO: 13ATGTTTGGTGTACAAGACACCCCGCAACATATATGCTACGAGCCTCAGCAGCGTAAGGTCAGTGAGAGAACATCACGTAACAGATCTCGTTCTAAATCACTGGACCCGGACAGCTTGCGCGAGAAAGGAAAGAAGACGCAACACCGTGAGGCGGATTGTTTCTTCGGTGAAGACAACCGGATGGAAAACTCCTACTCTGCGCAAATGTACATTGACGAGTTCGACCCTGTACACTATTACCAAACCTATTATTCCTCAGGGAAGGGCGGCATTGCTCGTGAGTGGACAGATTTTGCTTTGCAAAACTTGCATGAAACGTTCGGGCCTGGCGGGGTTAAAGGTGACATTCTTATTGACTTCGGTGCTGGGCCGACAATATATCAGCTTCTGAGCGCATGTGAGGTTTTCAATAGCATTATTACATCCGACTTTCTTGAGCAAAACCGCGAGCAACTTGAGAAATGGCTTCGAAAGGACCCGGACGCCCTTGACTGGTCCCATTTCACGAAGTACGTTTGCGAGCTCGAAGGCAACCGGGACAACTGGGAAAAGAAAGAGGAAACCCTGCGCCGAAAGGTTACCAAGGTGCTTAAATGTGACGCACTGGCCGAGAAGCCTTTCGACGACGTGCCAATGCCAGAGGCTGACTGTCTGATCTCATGCCTGTGTTTAGAGAACCCTTGTCAAGACCAGGAAGCTTACATTAACATATTGAAGAAGTTAAAAGAGCTCTTGAAACCGGGCGGCCACATCATTATACAGTCCATATTGAACTGCTCGTATTACCATATTGGCAATAGCTGCTTCTCACATTTGTCGTTAAGCAAGGACGACGTGGAGAAATCGTTTAAGGAAGCTGGCTACGAAATCGTCAAATTGAAGGTTCTTCCACGCTCAGTTATGTCGGAAATGGAAATCAGCGACTCAAATGGCTACTACTTCATCCACGCTCGGAAACCGCAAAAGGAGTAA SEQ. ID NO: 14MFGVQDTPQHICYEPQQRKVSERTSRNRSRSKSLDPDSLREKGKKTQHREADCFFGEDNRMENSYSAQMYIDEFDPVHYYQTYYSSGKGGIAREWTDFALQNLHETFGPGGVKGDILIDFGAGPTIYQLLSACEVFNSIITSDFLEQNREQLEKWLRKDPDALDWSHFTKYVCELEGNRDNWEKKEETLRRKVTKVLKCDALAEKPFDDVPMPEADCLISCLCLENPCQDQEAYINILKKLKELLKPGGHIIIQSILNCSYYHIGNSCFSHLSLSKDDVEKSFKEAGYEIVKLKVLPRSVMSEMEISDSNGYYFIHARKPQKE

EXAMPLES Example 1—Chemical Synthesis of a First, a Second and a ThirdAminated Psilocybin Derivative

Referring to FIG. 9D, shown therein is a 7-step synthesis for two4-O-methyl-psilocybin derivatives respectively aminated at C₅ and C₇from 4-methoxyindole.

The first step involved the regioselective 3-nitrovinylation of4-methoxyindole (9D-1). Under argon, 4-methoxy indole (9D-1) (4000 mg,27.18 mmol, 1.00 eq) and 1-(dimethylamino)-2-nitroethylene (3472 mg,29.90 mmol, 1.10 eq) were dissolved in trifluoroacetic acid (20.8 mL,272 mmol, 10.0 eq) and allowed to stir at room temperature for 3 hoursuntil complete as determined by TLC (1:1 ethyl acetate-hexanes). Thedark red solution was diluted with ethyl acetate (100 mL) and carefullypoured over saturated sodium bicarbonate solution (200 mL). Thisbiphasic mixture was then separated, and the aqueous phase extractedwith ethyl acetate (4×100 mL). The combined organic extracts were washedwith brine, dried with MgSO₄, and concentrated under reduced pressure toyield the crude product 9D-2 (5600 mg) that was used directly withoutany further purification.

The second step involved the conjugated reduction of the alkenefunctionality of compound 9D-2. To a solution of crude compound 9D-2(5600 mg) in ethanol (50 mL) and THE (50 mL) under ambient air was addedsodium borohydride (5141 mg, 136 mmol) in small portions, waiting foreffervescence to decrease between additions. This mixture was allowed tostir at room temperature for 18 hours at which point reaction wascomplete as determined by TLC (1:1 ethyl acetate-hexanes). The reactionwas quenched by pouring over ice-water (200 ml) and extracted with DCM(4×100 mL). The combined organic extracts were dried with MgSO₄ andconcentrated under reduced pressure to yield the crude product as abrown solid. Purification by column chromatography on silica gel using a10% to 50% ethyl acetate-hexanes gradient to yield compound 9D-3 as awhite solid (1950 mg, 8.94 mg, 33% over 2 steps). ¹H NMR (400 MHz,CDCl₃): δ (ppm)=8.02 (s, 1H), 7.15 (t, J=8.0 Hz, 1H), 7.00 (dd, J=8.3,0.7 Hz, 1H), 6.94 (dd, J=2.5, 1.0 Hz, 1H), 6.55 (dd, J=7.8, 0.7 Hz, 1H),4.76 (t, J=7.2 Hz, 2H), 3.97 (s, 3H), 3.57 (td, J=7.2, 0.8 Hz, 2H).

The third step involved the reduction of the nitro functionality ofcompound 3. Under argon in a flame-dried flask compound 9D-3 (800 mg,3.63 mmol, 1.00 eq) was dissolved in anhydrous THE (20 mL) and cooled to0° C. To this solution was added 1 M lithium aluminum hydride in THE(18.2 mL, 18.2 mmol, 5.00 eq), causing a colour change to yellow. Thereaction mixture was heated to reflux for 2.5 hours, yielding amilky-white solution. After cooling to 0° C., the reaction was quenchedwith 10% water-THE (15 mL) and allowed to stir for 10 minutes. Theprecipitated white solids were filtered off and the filter-cake rinsedwith THE (10 mL), dichloromethane (10 mL) and methanol (10 mL). Theorganic filtrate was dried with MgSO₄ and concentrated under reducedpressure to yield compound 9D-4 as an off-white solid (395 mg, 2.08mmol, 57%). ¹H NMR (400 MHz, CDCl₃): δ (ppm)=8.12 (s, 1H), 7.09 (t,J=7.9 Hz, 1H), 6.97 (d, J=8.1 Hz, 1H), 6.89 (d, J=2.2 Hz, 1H), 6.50 (d,J=7.8 Hz, 1H), 3.92 (s, 3H), 3.02 (s, 4H).

The fourth step involved the full protection of the side-chain aminogroup and the N1 of compound 9D-4. Under argon in a flame-dried flask,compound 9D-4 (245 mg, 1.29 mmol, 1.00 eq) was dissolved in anhydrousacetonitrile (10 mL). To this solution, di-tert-butyl dicarbonate (2811mg, 12.9 mmol, 10.0 eq) and 4-dimethylaminopyridine (157 mg, 1.29 mmol,1.00 eq) was added, and the reaction mixture allowed to stir at roomtemperature for 20 hours. Water (20 mL) was added, and the mixture wasextracted with dichloromethane (4×30 mL). The combined organic extractswere washed with brine (25 mL), dried with MgSO₄, and concentrated undervacuum to yield the crude product as a dark red oil. Purification bycolumn chromatography on silica gel using an 8% to 15% ethylacetate-hexanes gradient yielded compound 9D-5 as an oily white solid(325 mg, 0.663 mmol, 51%). ¹H NMR (400 MHz, CDCl₃): δ (ppm)=7.74 (d,J=8.4 Hz, 1H), 7.23-7.13 (m, 2H), 6.63 (d, J=7.9 Hz, 1H), 3.91 (m, 5H),3.08 (ddd, J=7.5, 6.3, 1.0 Hz, 2H), 1.63 (s, 9H), 1.38 (s, 18H).

The fifth step involved the regioselective nitration of compound 9D-5. Aflame-dried round-bottom flask was charged with compound 9D-5 (325 mg,0.662 mmol, 1.00 eq), silver nitrate (124 mg, 0.729 mmol, 1.10 eq), anddry acetonitrile (2.5 mL), then cooled to 0° C. under argon atmosphere.Benzoyl chloride (102 mg, 0.729 mmol, 1.10 eq) was diluted with dryacetonitrile (0.5 mL) and added dropwise to the reaction mixture, whichwas then allowed to stir at 0° C. for 3 hours. The reaction mixture wasdiluted with ethyl acetate (10 mL) and the precipitated salts wereremoved via vacuum filtration and washed with ethyl acetate (5 mL). Theorganic filtrate was washed with water (3×20 mL) and saturated Na₂CO₃(20 mL), then dried with MgSO₄ and solvent removed in vacuo. The crudemixture was purified by column chromatography on silica gel using agradient of 5 to 15% ethyl acetate-hexanes to afford compounds 9D-6a (25mg, 0.047 mmol, 7%), 9D-6b (48 mg, 0.090 mmol, 14%), and 9D-6c (45 mg,0.078 mmol, 12%) in order of elution as yellow solids. Compound 9D-6a:¹H NMR (400 MHz, CDCl₃): δ (ppm)=7.62 (dd, J=8.5, 0.7 Hz, 1H), 7.41 (t,J=8.3 Hz, 1H), 7.41 (s, 1H), 6.70 (dd, J=8.1, 0.7 Hz, 1H), 4.07-4.02 (m,2H), 3.94 (s, 3H), 3.37-3.31 (m, 2H), 1.55 (s, 9H), 1.33 (s, 18H).Compound 9D-9D-6b: ¹H NMR (400 MHz, CDCl₃): δ (ppm)=8.00-7.94 (m, 1H),7.87 (d, J=9.0 Hz, 1H), 7.38 (s, 1H), 4.01 (s, 3H), 3.94 (t, J=7.6 Hz,2H), 3.10 (t, J=7.6 Hz, 2H), 1.65 (s, 9H), 1.41 (s, 18H). Compound9D-6c: ¹H NMR (400 MHz, CDCl₃): δ (ppm)=7.82 (d, J=8.8 Hz, 1H), 6.64 (d,J=8.8 Hz, 1H), 4.02 (s, 3H), 3.91 (dd, J=7.4, 6.4 Hz, 2H), 3.08 (td,J=6.8, 6.4, 1.0 Hz, 2H), 1.57 (s, 9H), 1.40 (s, 18H).

The sixth step involved the reduction of the nitro group. To avigorously stirring solution of compound 9D-6b (27 mg, 0.050 mmol, 1.0eq) in methanol (2 mL) was added 10% palladium on activated charcoal (23mg, 0.021 mmol, 0.50 eq) followed by ammonium formate (46 mg, 0.70 mmol,17 eq). The reaction mixture was allowed to stir at room temperature for2 hours until complete as determined by TLC (1:4 ethyl acetate-hexanes).The catalyst was removed and methanol was removed under vacuum. Theresidue was taken up in dichloromethane (10 mL), washed with brine (10mL), the organic phase dried with MgSO₄ and concentrated under reducedpressure to yield compound 9D-7 as a colourless film (22 mg, 0.044 mmol,88%). ¹H NMR (400 MHz, CDCl₃): δ (ppm)=7.72 (s, 1H), 7.26-7.18 (m, 1H),6.76 (d, J=8.7 Hz, 1H), 3.96-3.91 (m, 2H), 3.84 (s, 3H), 3.09-3.00 (m,2H), 1.62 (s, 9H), 1.43 (s, 18H).

Similarly, the nitro group of compound 9D-6c can also be reduced in ananalogous manner. To a vigorously stirring solution of compound 9D-6c(22 mg, 0.041 mmol, 1.0 eq) in methanol (2 mL) was added 10% palladiumon activated charcoal (22 mg, 0.021 mmol, 0.50 eq) followed by ammoniumformate (44 mg, 0.70 mmol, 17 eq). The reaction mixture was allowed tostir at room temperature for 2 hours and monitored by TLC (1:4 ethylacetate-hexanes). The catalyst was removed and methanol was removedunder vacuum. The residue was taken up in dichloromethane (10 mL),washed with brine (10 mL), the organic phase dried with MgSO₄ andconcentrated under reduced pressure to yield compound 9D-9 as a brownfilm (14 mg, 0.028 mmol, 67%). ¹H NMR (600 MHz, CDCl₃): δ (ppm)=7.13 (s,1H), 6.69 (d, J=8.3 Hz, 1H), 6.57 (dt, J=8.5, 1.0 Hz, 1H), 3.93-3.87 (m,2H), 3.85 (s, 3H), 3.04 (t, J=6.9 Hz, 2H), 1.61 (s, 9H), 1.40 (s, 18H).

The seventh step involved the removal of all protecting group. To asolution of 9D-7 (22 mg, 0.044 mmol, 1.0 eq) in dichloromethane (1.0 mL)and methanol (0.1 mL) was added trifluoroacetic acid (0.15 mL, 1.5 mmol,35 eq) dropwise. The reaction mixture was heated to 40° C. for 2 hours,then allowed to stir at room temperature for 18 hours. As the reactionwas incomplete, the reaction was heated for a further 20 hours at 40° C.The reaction mixture was concentrated under reduced pressure to yieldcompound 9D-8 (5 mg, 0.2 mmol, 56%). ¹H NMR (600 MHz, D₂O): δ (ppm)=7.24(d, J=8.5 Hz, 1H), 7.22 (s, 1H), 7.03 (d, J=8.5 Hz, 1H), 3.81 (s, 3H),3.23 (t, J=7.0 Hz, 2H), 3.10 (t, J=7.0 Hz, 2H).

It is noted that compound 9D-8 corresponds with an example compoundhaving chemical formula (IX):

set forth herein.

Similarly, compound 9D-9 can also be deprotected in an analogous manner.To a solution of 9D-9 (14 mg, 0.028 mmol, 1.0 eq) in dichloromethane(1.0 mL) and methanol (0.1 mL) was added trifluoroacetic acid (0.064 mL,0.83 mmol, 30 eq). The reaction mixture was heated to 40° C. for 20hours. The reaction mixture was concentrated under reduced pressure toyield compound 9D-10 (4 mg, 0.2 mmol, 70%). ¹H NMR (600 MHz, D₂O): δ(ppm)=7.25 (s, 1H), 7.18 (d, J=8.3 Hz, 1H), 6.66 (d, J=8.3 Hz, 1H), 3.96(s, 3H), 3.30 (t, J=6.8 Hz, 2H), 3.21 (t, J=6.9 Hz, 2H).

It is noted that compound 9D-10 corresponds with an example compoundhaving chemical formula (XII):

set forth herein.

Assessment of Cell Viability Upon Treatment of Aminated PsilocybinDerivatives

To establish suitable ligand concentrations for competitive bindingassays, PrestoBlue assays were first performed. The PrestoBlue assaymeasures cell metabolic activity based on tetrazolium salt formation,and is a preferred method for routine cell viability assays (Terrasso etal., 2017, J Pharmacol Toxicol Methods 83: 72). Results of these assayswere conducted using both control ligands (e.g., psilocybin, psilocin,DMT) and novel derivatives, in part as a pre-screen for any remarkabletoxic effects on cell cultures up to concentrations of 1 mM. A knowncellular toxin (Triton X-100, Pyrgiotakis G. et al., 2009, Ann. Biomed.Eng. 37: 1464-1473) was included as a general marker of toxicity.Drug-induced changes in cell health within simple in vitro systems suchas the HepG2 cell line are commonly adopted as first-line screeningapproaches in the pharmaceutical industry (Weaver et al, 2017, ExpertOpin Drug Metab Toxicol 13: 767). HepG2 is a human hepatoma that is mostcommonly used in drug metabolism and hepatotoxicity studies (Donato etal., 2015, Methods Mol Biol 1250: 77). Herein, HepG2 cells were culturedusing standard procedures using the manufacture's protocols (ATCC,HB-8065). Briefly, cells were cultured in Eagle's minimum essentialmedium supplemented with 10% fetal bovine serum and grown at 37° C. inthe presence of 5% CO₂. To test the various compounds with the cellline, cells were seeded in a clear 96-well culture plate at 20,000 cellsper well. After allowing cells to attach and grow for 24 hours,compounds were added at 1 μM, 10 μM, 100 μM, and 1 mM. Methanol was usedas vehicle, at concentrations 0.001, 0.01, 0.1, and 1%. As a positivecontrol for toxicity, TritonX concentrations used were 0.0001, 0.001,0.01 and 0.1%. Cells were incubated with compounds for 48 hours beforeaccessing cell viability with the PrestoBlue assay following themanufacture's protocol (ThermoFisher Scientific, P50200). PrestoBluereagent was added to cells and allowed to incubate for 1 hour beforereading. Absorbance readings were performed at 570 nm with the referenceat 600 nm on a SpectraMax iD3 plate reader. Non-treated cells wereassigned 100% viability. Bar graphs show the mean +/−SD, n=3.Significance was determined by 2-way ANOVA followed by Dunnett'smultiple comparison test and is indicated by *** (P<0.0001),**(P<0.001), *(P<0.005). Data acquired for the derivative havingchemical formula (IX) is displayed as “IX” on the x-axis of FIG. 11A.).Data acquired for the derivative having chemical formula (XII) isdisplayed as “XII” on the x-axis of FIG. 11B.

Radioligand Receptor Binding Assays.

Evaluation of drug binding is an essential step to characterization ofall drug-target interactions (Fang 2012, Exp Opin Drug Discov 7:969).The binding affinity of a drug to a target is traditionally viewed as anacceptable surrogate of its in vivo efficacy (NGAez et al., 2012, DrugDisc Today 17: 10). Competition assays, also called displacement ormodulation binding assays, are a common approach to measure activity ofa ligand at a target receptor (Flanagan 2016, Methods Cell Biol 132:191). In these assays, standard radioligands acting either as agonistsor antagonists are ascribed to specific receptors. In the case of Gprotein-coupled receptor 5-HT_(2A), [³H]ketanserin is a well-establishedantagonist used routinely in competition assays to evaluate competitiveactivity of novel drug candidates at the 5-HT_(2A) receptor (Maguire etal., 2012, Methods Mol Biol 897: 31). Thus, to evaluate activity ofnovel psilocybin derivatives at the 5-HT_(2A) receptor, competitionassays using [³H]ketanserin were employed as follows. SPA beads(RPNQ0010), [³H] ketanserin (NET1233025UC), membranes containing5-HT_(2A) (ES-313-M400UA), and isoplate-96 microplate (6005040) were allpurchased from PerkinElmer. Radioactive binding assays were carried outusing Scintillation Proximity Assay (SPA). For saturation bindingassays, mixtures of 10 ug of membrane containing 5-HT_(2A) receptor waspre-coupled to 1 mg of SPA beads at room temperature in a tube rotatorfor 1 hour in binding buffer (50 mM Tris-HCl pH7.4, 4 mM CaCl₂, 1 mMascorbic acid, 10 μM pargyline HCl). After pre-coupling, the beads andmembrane were aliquoted in an isoplate-96 microplate with increasingamounts of [³H]ketanserin (0.1525 nM to 5 nM) and incubated for twohours at room temperature in the dark with shaking. After incubation,the samples were read on a MicroBeta 2 Microplate Counter (PerkinElmer). Determination of non-specific binding was carried out in thepresence of 20 μM of spiperone (S7395-250MG, Sigma). Equilibrium bindingconstants for ketanserin (K_(d)) were determined from saturation bindingcurves using the ‘one-site saturation binding analysis’ method ofGraphPad PRISM software (Version 9.2.0). Competition binding assays wereperformed using fixed (1 nM) [³H]ketanserin and different concentrationsof tryptophan (3 nM to 1 mM), psilocin (30 pM to 10 μM) or unlabeledtest compound (3 nM to 1 mM) similar to the saturation binding assay.K_(i) values were calculated from the competition displacement datausing the competitive binding analysis from GraphPad PRISM software.Tryptophan was included as a negative control as it has no activity atthe 5-HT_(2A) receptor. In contrast, psilocin was used as a positivecontrol since it has established binding activity at the 5-HT_(2A)receptor (Kim et al., 2020, Cell 182: 1574). FIG. 11D depicts thesaturation binding curves for [³H]ketanserin at the 5-HT_(2A) receptor.Panel 1 shows the specific saturation ligand binding of [³H]ketanserin(from 0.1525 nM to 5 nM) to membranes containing 5-HT_(2A) receptor,which was obtained after subtracting non-specific binding values (shownin Panel 2). Specific binding in counts per minute (cpm) was calculatedby subtracting non-specific binding from total binding. Specific binding(pmol/mg) was calculated from pmol of [³H]ketanserin bound per mg ofprotein in the assay. The K_(d) was calculated by fitting the data withthe one-site binding model of PRISM software (version 9.2.0). FIG. 11E(top panel) shows the competition binding curve for psilocin as apositive control (binding). FIG. 11E (bottom panel) shows thecompetition binding curve for tryptophan as a negative control (nobinding). FIG. 11F shows competition binding curve for compound withformula (IX), designated “IX” in the figure. FIG. 11G shows competitionbinding curve for compound with formula (XII), designated “XII” in thefigure.

Cell Lines and Control Ligands Used to Assess Activity at 5-HT_(1A).

CHO-K1/Gα₁₅ (GenScript, M00257) (−5-HT_(1A)) and CHO-K1/5-HT_(1A)/Gα₁₅(GenScript, M00330) (+5-HT_(1A)) cells lines were used. Briefly,CHO-K1/Gα₁₅ is a control cell line that constitutively expresses Gα₁₅which is a promiscuous G_(q) protein. This control cell line lacks anytransgene encoding 5-HT_(1A) receptors, but still responds to forskolin;thus, cAMP response to forskolin should be the same regardless ofwhether or not 5-HT_(1A) agonists are present. Conversely,CHO-K1/5-HT_(1A)/Gα₁₅ cells stably express 5-HT_(1A) receptor in theCHO-K1 host background. Notably, Gα₁₅ is a promiscuous G protein knownto induce calcium flux response, present in both control and 5-HT_(1A)cell lines. In +5-HT_(1A) cells, Gα₁₅ may be recruited in place ofG_(αi/o), which could theoretically dampen cAMP response (Rojas andFiedler 2016, Front Cell Neurosci 10: 272). Thus, we included two known5-HT_(1A) agonists, DMT (Cameron and Olson 2018, ACS Chem Neurosci 9:2344) and serotonin (Rojas and Fiedler 2016, Front Cell Neurosci 10:272) as positive controls to ensure sufficient cAMP response wasobserved, thereby indicating measurable recruitment of G_(αi/o) proteinto activated 5-HT_(1A) receptors. In contrast, tryptophan is not knownto activate, or modulate in any way, 5-HT_(1A) receptors, and was thusused as a negative control. Cells were maintained in complete growthmedia as recommended by supplier (GenScript) which is constituted asfollows: Ham's F12 Nutrient mix (HAM's F12, GIBCO #11765-047) with 10%fetal bovine serum (FBS) (Thermo Scientific #12483020), 200 μg/ml zeocin(Thermo Scientific #R25005) and/or 100 μg/ml hygromycin (ThermoScientific #10687010). The cells were cultured in a humidified incubatorwith 37° C. and 5% CO₂. Cells maintenance was carried out as recommendedby the cell supplier. Briefly, vials with cells were removed from theliquid nitrogen and thawed quickly in 37° C. water bath. Just before thecells were completely thawed the vial's outside was decontaminated by70% ethanol spray. The cell suspension was then retrieved from the vialand added to warm (37° C.) complete growth media, and centrifuged at1,000 rpm for 5 minutes. The supernatant was discarded, and the cellpellet was then resuspended in another 10 ml of complete growth media,and added to the 10 cm cell culture dish (Greiner Bio-One #664160). Themedia was changed every third day until the cells were about 90%confluent. The ˜90% confluent cells were then split 10:1 for maintenanceor used for experiment.

Evaluation of 5-HT_(1A) Receptor Modulation

As 5-HT_(1A) activation inhibits cAMP formation, the ability of testmolecules to modulate 5-HT_(1A) response was measured via changes in thelevels of cAMP produced due to application of 4 μM forskolin. The changein intracellular cAMP levels due to the treatment of novel molecules wasmeasured using cAMP-Glo Assay kit (Promega #V1501). Briefly, +5-HT_(1A)cells were seeded on 1-6 columns and base −5-HT_(1A) cells were seededon columns 7-12 of the white walled clear bottom 96-well plate (Corning,#3903). Both cells were seeded at the density of 30,000 cells/well in100 μl complete growth media and cultured 24 hrs in humidified incubatorat 37° C. and 5% CO₂. On the experiment day, the media of cells wasreplaced with serum/antibiotic free culture media. Then the cells weretreated for 20 minutes with test molecules dissolved in induction medium(serum/antibiotic free culture media containing 4 μM forskolin, 500 μMIBMX (isobutyl-1-methylxanthine, Sigma-Aldrich, Cat. #17018) and 100 μM(RO 20-1724, Sigma-Aldrich, Cat. #B8279)). Forskolin induced cAMPformation whereas IBMX and RO 20-1724 inhibited the degradation of cAMP.PKA was added to the lysate, mixed, and subsequently the substrate ofthe PKA was added. PKA was activated by cAMP, and the amount of ATPconsumed due to PKA phosphorylation directly corresponded to cAMP levelsin the lysate. Reduced ATP caused reduced conversion of luciferin tooxyluciferin, conferring diminished luminescence as the result of5-HT_(1A) activation. Conversely, enhanced luminescence was expected incases where 5-HT_(1A) receptor modulation—imparted by a testmolecule—caused downstream increases in ATP, thus imparting enhancedconversion of luciferin to oxyluciferin. FIG. 11I shows increasedluminescence resulting from decreased dosages of forskolin (anddecreased cAMP) in +5HT_(1A) cell culture. FIG. 11J illustrates reducedluminescence (i.e., increased cAMP) in the presence of fixed (4 μM)forskolin as dosages of DMT decrease, revealing 5-HT_(1A) activity ofDMT. FIG. 11K illustrates no trend in luminescence (i.e., no trend incAMP levels) in the presence of fixed (4 μM) forskolin, as dosages oftryptophan decrease, revealing a lack of 5-HT_(1A) modulation fortryptophan. FIG. 11L illustrates increased % cAMP levels in the presenceof fixed (4 μM) forskolin as dosages of serotonin decrease, revealing5-HT_(1A) binding activity of serotonin in +5HT_(1A) cell cultures.Conversely, this trend of increasing % cAMP levels with decreasingserotonin is not observed in −5HT_(1A) cell cultures.

FIG. 11M illustrates increased % cAMP levels in the presence of fixed (4μM) forskolin as dosages of compound (IX) decrease, revealing 5-HT_(1A)binding activity of compound (IX) in +5HT_(1A) cell cultures.Conversely, this trend of increasing % cAMP levels with decreasingcompound (IX) is not observed in −5HT_(1A) cell cultures. Note thatcompound (IX) is shown simply as (IX) along the x-axis. For FIGS.11I-11M, error bars represent results of three experiments (n=3).

Example 2—Biochemical Synthesis of a Fourth Aminated PsilocybinDerivative

E. coli strain Ec-1 was constructed as follows. For plasmid cloning,Top10 or XL1-blue strains were used depending on antibiotic markers.Standard LB media was used for culturing. For gene expression andfeeding experiments, the parent host strain employed was BL21 (DE3). Theplasmid pETM6-H10-TmTrpB-2F3-V5-BaTDC-FLAG was created by first cloningthe in-frame, C-terminally V5-tagged (SEQ. ID NO: 6, SEQ. ID NO: 7)TmTrpB-2F3 (SEQ. ID NO: 8, SEQ. ID NO: 9) into the NdeI/XhoI site ofpETM6-H10 (SEQ. ID NO: 10) to create pETM6-H₁₀-TmTrpB-2F3-V5. Thisintermediate plasmid was digested with SpeI and SalI, and in-frame,C-terminally FLAG tagged (SEQ. ID NO: 2, SEQ. ID NO: 3) BaTDC (SEQ. IDNO: 11, SEQ. ID NO: 12) was cloned into the site with XbaI and SalI,nullifying the SpeI restriction site. In this setup, the T7 polymerasewas able to drive the expression of the polycistronic DNA containingboth TmTrpB-2F3 and BaTDC. The target plasmidpETM6-H₁₀-TmTrpB-2F3-V5-BaTDC-FLAG was transformed into BL21 (DE3)cells, and ampicillin was used to select for the correct clonescontaining the plasmid. Scaled-up culturing of engineered E. coli wasconducted as follows: seed cultures were inoculated in AMM (Jones et al.2015, Sci Rep. 5: 11301) medium overnight. The overnight culture wasthen divided into two flasks containing 500 mL each of AMM mediumadditionally containing 0.5% (w/v) serine, 1M IPTG, 50 ug/L ampicillin,and 100 mg/L aminated indole feedstock (5,7-dimethyl-1H-indol-4-ylamine;1clickchemistry, www.1 clickchemistry.com) for conversion by Ec-1.Cultures were grown for 24 h. Cultures were then centrifuged (10,000 g×5minutes) to remove cellular content, and culture broth containingsecreted derivative was stored at −80° C. until further processing.

Analysis and Purification,

Analysis was carried out using high-resolution LC-HESI-LTQ-Orbitrap-XLMS (Thermo Fisher Scientific), employing a modified version of a methoddescribed previously (Chang et al., 2015, Plant Physiol. 169:1127-1140), with the exception that liquid chromatography was carriedout using an UltiMate 3000 HPLC (Thermo Fisher Scientific) equipped witha Poroshell 120 SB-C18 column (Agilent Technologies) instead of anAccela HPLC system (Thermo Fisher Scientific) equipped with a Zorbax C18column (Agilent Technologies). Briefly, 100 microliters of culture mediawere were dried and resuspended in 100 microliters of DMSO. One tenth(10 microliters) of this suspension was injected at a flow rate of 0.5mL/min and a gradient of solvent A (water with 0.1% of formic acid) andsolvent B (ACN with 0.1% formic acid) as follows: 100% to 0% (v/v)solvent A over 5 min; isocratic at 0% (v/v) for 1 min; 0% to 100% (v/v)over 0.1 min; and isocratic at 100% (v/v) for 1.9 min. Total run timewas 8 minutes. Heated ESI source and interface conditions were operatedin positive ion mode as follows: vaporizer temperature, 400° C.; sourcevoltage, 3 kV; sheath gas, 60 au, auxiliary gas, 20 au; capillarytemperature, 380° C.; capillary voltage, 6 V; tube lens, 45 V.Instrumentation was performed as a single, HR scan event using Orbitrapdetection of m/z in the range of 100-500 m/z. Ion injection time was 300ms with scan time of 1 s. External and internal calibration proceduresensured <2 ppm error to facilitate elemental formulae predictions.Singly protonated product with exact m/z and expected elemental formulamatching the singly protonated form of3-(2-aminoethyl)-5,7-dimethyl-1H-indol-4-amine, having chemical formula(VI):

eluted at 2.9 minutes (EIC, see: FIG. 12A). Although peak splitting canbe minimized through the use of DMSO as injection solvent (Kaufman andJegle 2005, Agilent Technologies Technical Bulletin 5989-2485EN), thisphenomenon persisted owing to ion pairing effects between matrixcomponents (Tarafder et al. 2010, J Chromatogr A 1217:7065-7073).

As per standard procedures (Menéndez-Perdomo et al. 2021, Mass Spectrom56: 34683) further analysis using high energy collisions (HCD) wasachieved in a dedicated, post-LTQ, nitrogen collision cell.Orbitrap-based, HR fragment detection was employed (normalized collisionenergy, NCE 35), enabling opportunity to assign elemental formulae tosubsequent diagnostic ion species characteristic of the targetedaminated psilocybin derivative with formula (VI) as follows (FIG. 12B,Table 1) (Servillo L et al., 2013, J. Agric. Chem. 61: 5156-5162).

TABLE I % Relative Ionic Empirical m/z abundance species formula204.1131 100 [M + H]⁺ C₁₂H₁₅N₃ 187.1229 55 [M + H − NH₂]⁺ C₁₂H₁₆N₂202.4920 21 186.1106 19 189.0976 6.0 200.0973 5.0 162.1027 4.1 150.68543.8 92.8785 3.8 61.0962 3.8

Example 3—Biochemical Synthesis of a Fifth Aminated PsilocybinDerivative

Escherichia coli strain Ec-1 was used to biosynthesize aminatedpsilocybin derivative with formula (XIV) from aminated indole feedstock.The construction of Ec-1 is described in Example 2. Scaled-up culturingand material storage of engineered E. coli was conducted as described inExample 2, except that 6-methyl-1H-indol-4-ylamine (Combi-Blocks,www.combi-blocks.com) was used in place of5,7-dimethyl-1H-indol-4-ylamine. To assess product, high-resolutionLC-HESI-LTQ-Orbitrap-XL MS analysis was conducted as described inExample 2. Singly protonated product with exact m/z and expectedelemental formula matching the singly protonated form of3-(2-aminoethyl)-6-methyl-1H-indol-4-amine having chemical formula(XIV):

eluted at 2.4 minutes (EIC, see: FIG. 13A). As per standard procedures(Menéndez-Perdomo et al. 2021, Mass Spectrom 56: 34683) high energycollisions (HCD) were achieved in a dedicated, post-LTQ, nitrogencollision cell. Orbitrap-based, HR fragment detection was employed(normalized collision energy, NCE 35), enabling opportunity to assignelemental formulae to subsequent diagnostic ion species characteristicof a compound of formula (XIV), as follows (FIG. 13B, Table II)(Servillo L. et al., 2013, J. Agric. Chem. 61: 5156-5162),

TABLE II % Relative Ionic m/z abundance species Δ ppm 173.10707 100 [M +H − NH₂]⁺ 1.44 130.06489 20 144.08057 16 190.13373 2.7 [M + H]⁺ 1.00161.10715 2.6 118.06485 1.9 202.35010 1.5 160.07551 1.0

Example 4—Biochemical Synthesis of a Sixth Aminated PsilocybinDerivative

Escherichia coli strain Ec-2 was used to biosynthesize aminatedpsilocybin derivative, where the amino group terminating the 2-carbonaliphatic side chain was conjugated to an acetyl group. Ec-2 wasconstructed using the same method as for Ec-1 (see: Example 2), exceptthat an additional plasmid was assembled and transformed into cellsalong with pETM6-H₁₀-TmTrpB-2F3-V5-BaTDC-FLAG. This additional plasmidencoded a promiscuous and efficient Streptomyces griseofuscusN-acetyltransferase enzyme named PsmF (SEQ. ID NO: 5). This additionalplasmid was assembled as follows: from plasmid pCDM4 (SEQ. ID NO: 1),the plasmid pCDM4-PsmF-FLAG was created by inserting an in-frame,C-terminally FLAG-tagged (SEQ. ID NO: 2, SEQ. ID NO: 3) PsmF gene (SEQ.ID NO: 4, SEQ. ID NO: 5) into the NdeI/XhoI site of pCDM4. The twotarget plasmids pCDM4-PsmF-FLAG and pETM6-H10-TmTrpB-2F3-V5-BaTDC-FLAGwere transformed into BL21 (DE3) cells, and antibiotics ampicillin plusstreptomycin were used to select for the correct clones containing bothplasmids. Scaled up culturing, analysis, purification, toxicology, andpharmacological testing were performed as described in Example 2, exceptthat 1H-indol-7-ylamine (Combi-Blocks, www.combi-blocks.com) was used inplace of 5,7-dimethyl-1H-indol-4-ylamine. Purification of the targetproduct was achieved as follows: to 0.75 L of E. coli culture, 10M NaOHsolution was added until the pH reached ˜7. The culture was thenextracted by ethyl acetate (4×500 ml). The organic layer was dried overNa₂SO₄, followed by concentration under reduced pressure. The residuewas purified by flash chromatography on silica gelmethanol-dichloromethane (2->4%) as eluent, to give the compound aslight yellow solid (15 mg). NMR and HRMS data were as follows: ¹H NMR(400 MHz, CD₃OD): δ=1.91 (s, 3H), 2.22 (s, 3H), 2.93 (t, J=6.9 Hz, 2H),3.46 (t, J=7.2 Hz, 2H), 6.99 (t, J=7.8 Hz, 1H), 7.10 (m, 2H), 7.44 (dd,J=7.9, 1.0 Hz, 1H). Selective ¹³C NMR (100 MHz, CD₃OD): δ=21.2, 21.8,24.7, 40.0, 112.5, 115.2, 115.8, 118.3, 122.1, 122.3, 129.4, 170.5,171.8. HRMS (ESI) m/z: calcd. for C₁₄H₁₇N₃O₂ [M+H]⁺ 260.1394, found260.1392. Purity was assessed at 95%. This characterization confirmed astructure corresponding to compound (XIII):

Assessment of Cell Viability Upon Treatment of Aldehyde PsilocybinDerivative

Cell viability was assessed as described for Example 1, except thecompound with formula (XIII) was evaluated in place of the compoundswith formulae (IX) and (XII). FIG. 11C shows PrestoBlue assay resultsfor compound with formula (XIII), depicted on the x-axis as “XIII”.

Radioligand Receptor Binding Assays.

Activity at 5-HT_(2A) receptor was assessed as described for Example 1,except the compound with formula (XIII) was evaluated in place of thecompounds with formulae (IX) and (XII). FIG. 11H shows radioligandcompetition assay results for compound with formula (XIII), depicted onthe x-axis simply as “XIII”.

Example 5—Biochemical Synthesis of a Seventh Aminated PsilocybinDerivative

Escherichia coli strain Ec-2 was used to biosynthesize aminatedpsilocybin derivative with formula (VII) from aminated indole feedstock.The construction of Ec-2 is described in Example 4. Scaled-up culturingand material storage of engineered E. coli was conducted as described inExample 2. To assess product, high-resolution LC-HESI-LTQ-Orbitrap-XL MSanalysis was conducted as described in Example 2. Singly protonatedproduct with exact m/z and expected elemental formula matching thesingly protonated form ofN-[2-(4-amino-5,7-dimethyl-1H-indol-3-yl)ethyl]acetamide having chemicalformula (VII):

eluted at 2.9 minutes (EIC, see: FIG. 14A). Although peak splitting canbe minimized through the use of DMSO as injection solvent (Kaufman andJegle 2005, Agilent Technologies Technical Bulletin 5989-2485EN), thisphenomenon persisted owing to ion pairing effects between matrixcomponents (Tarafder et al 2010, J Chromatogr A 1217:7065-7073).

As per standard procedures (Menéndez-Perdomo et al. 2021, Mass Spectrom56: 34683) high energy collisions (HCD) were achieved in a dedicated,post-LTQ, nitrogen collision cell. Orbitrap-based, HR fragment detectionwas employed (normalized collision energy, NCE 35), enabling opportunityto assign elemental formulae to subsequent diagnostic ion speciescharacteristic of a compound of formula (VII), as follows (FIG. 14B,Table 111) (Servillo L. et al, 2013, J. Agric. Chen. 61: 5156-5162).

TABLE III % Relative Ionic Empirical m/z Abundance species formula187.1228 100 [M + H − N-acetyl]⁺ C₁₂H₁₅N₂ 246.1601 23 [M + H]⁺ C₁₄H₂₀N₃O229.1335 16 [M + H − NH₂]⁺ C₁₄H₁₇N₂O 187.1257 1.2 203.2231 1.0 217.13340.9 187.1191 0.6 93.0529 0.6 66.1240 0.6

Example 6—Biochemical Synthesis of an Eighth Aminated PsilocybinDerivative

Escherichia coli strain Ec-2 was used to biosynthesize aminatedpsilocybin derivative with formula (IV) from aminated indole feedstock.The construction of Ec-2 is described in Example 4. Scaled-up culturingand material storage of engineered E. coli was conducted as described inExample 2, except that 1H-indol-6-ylamine was used in place of5,7-dimethyl-1H-indol-4-ylamine. To assess product, high-resolutionLC-HESI-LTQ-Orbitrap-XL MS analysis was conducted as described inExample 2. Singly protonated product with exact m/z and expectedelemental formula matching the singly protonated form ofN-[2-(6-amino-1H-indol-3-yl)ethyl]acetamide having chemical formula(IV):

eluted at 2.3 minutes (EIC, see: FIG. 15A).

As per standard procedures (Menéndez-Perdomo et al. 2021, Mass Spectrom56: 34683) high energy collisions (HCD) were achieved in a dedicated,post-LTQ, nitrogen collision cell. Orbitrap-based, HR fragment detectionwas employed (normalized collision energy, NCE 35), enabling opportunityto assign elemental formulae to subsequent diagnostic ion speciescharacteristic of a compound of formula (IV), as follows (FIG. 15B,Table IV) (Servillo L. et al., 2013, J. Agric. Chem. 61: 5156-5162).

TABLE IV % Relative Ionic Empirical m/z Abundance Species Formula159.0916 100 [M + H − NH₂—C₂H₃O]⁺ C₁₀H₁₁N₂ 201.1022 12 [M + H − NH₂]⁺C₁₂H₁₃N₂O 218.1288 5.1 [M + H]⁺ C₁₂H₁₆N₃O 202.4931 1.2 159.0946 1.1160.4383 0.5 167.4664 0.3 83.3505 0.3 56.9868 0.3

Example 7—Biochemical Synthesis of a Ninth Aminated PsilocybinDerivative

Escherichia coli strain Ec-2 was used to biosynthesize aminatedpsilocybin derivative with formula (III) from aminated indole feedstock.The construction of Ec-2 is described in Example 4. Scaled-up culturingand material storage of engineered E. coli was conducted as described inExample 2, except that 1H-indol-4-ylamine (Combi-Blocks;www.combi-blocks.com) was used in place of5,7-dimethyl-1H-indol-4-ylamine. To assess product, high-resolutionLC-HESI-LTQ-Orbitrap-XL MS analysis was conducted as described inExample 2. Singly protonated product with exact m/z and expectedelemental formula matching the singly protonated form ofN-[2-(4-amino-1H-indol-3-yl)ethyl]acetamide having chemical formula(III):

eluted at 2.5 minutes (EIC, see: FIG. 16A). Although peak splitting canbe minimized through the use of DMSO as injection solvent (Kaufman andJegle 2005, Agilent Technologies Technical Bulletin 5989-2485EN), thisphenomenon persisted owing to ion pairing effects between matrixcomponents (Tarafder et al. 2010, J Chromatogr A 1217:7065-7073).

As per standard procedures (Menéndez-Perdomo et al. 2021, Mass Spectrom56: 34683) high energy collisions (HCD) were achieved in a dedicated,post-LTQ, nitrogen collision cell. Orbitrap-based, HR fragment detectionwas employed (normalized collision energy, NCE 35), enabling opportunityto assign elemental formulae to subsequent diagnostic ion speciescharacteristic of a compound of formula (III), as follows (FIG. 16B,Table V) (Servillo L. et al., 2013, J. Agric. Chem. 61: 5156-5162).

TABLE V % Relative Ionic Empirical m/z Abundance Species Formula159.0915 100 218.1288 56 [M + H]⁺ C₁₂H₁₆N₃O 201.1022 23 C₁₂H₁₃N₂O202.4901 2.1 189.1022 1.0 C₁₁H₁₃N₂O 176.1182 0.6 C₁₀H₁₄N₃ 200.1049 0.6122.0354 0.5 183.0916 0.4 61.0965 0.4

Example 8—Biochemical Synthesis of a Tenth Aminated PsilocybinDerivative

Escherichia coli strain Ec-2 was used to biosynthesize aminatedpsilocybin derivative with formula (XV) from aminated indole feedstock.The construction of Ec-2 is described in Example 4. Scaled-up culturingand material storage of engineered E. coli was conducted as described inExample 2, except that 6-methyl-1H-indol-4-ylamine (BLDPharm;www.bldpharm.com) was used in place of 5,7-dimethyl-1H-indol-4-ylamine.To assess product, high-resolution LC-HESI-LTQ-Orbitrap-XL MS analysiswas conducted as described in Example 2. Singly protonated product withexact m/z and expected elemental formula matching the singly protonatedform of N-[2-(4-amino-6-methyl-1H-indol-3-yl)ethyl]acetamide havingchemical formula (XV):

eluted at 2.9 minutes (EIC, see: 17A). As per standard procedures(Menéndez-Perdomo et al. 2021, Mass Spectrom 56: 34683) high energycollisions (HCD) were achieved in a dedicated, post-LTQ, nitrogencollision cell. Orbitrap-based, HR fragment detection was employed(normalized collision energy, NCE 35), enabling opportunity to assignelemental formulae to subsequent diagnostic ion species characteristicof a compound of formula (XV), as follows (17B, Table VI) (Servillo L.et al., 2013, J. Agric. Chem. 61: 5156-5162).

TABLE VI % Relative Ionic m/z abundance species Δ ppm 173.10712 100 [M +H − C₂H₅NO]⁺ 1.55 232.14436 64 [M + H]⁺ 0.30 215.11772 14 201.98703 1.3203.11772 1.1 66.12433 0.8 95.31106 0.8 78.58906 0.7

Example 9—Biochemical Synthesis of an Eleventh Aminated PsilocybinDerivative

Escherichia coli strain Ec-2 was used to biosynthesize aminatedpsilocybin derivative with formula (XVI) from aminated indole feedstock.The construction of Ec-2 is described in Example 4. Scaled-up culturingand material storage of engineered E. coli was conducted as described inExample 2, except that 7-methyl-1H-indol-5-ylamine (BLDPharm;www.bldpharm.com) was used in place of 5,7-dimethyl-1H-indol-4-ylamine.To assess product, high-resolution LC-HESI-LTQ-Orbitrap-XL MS analysiswas conducted as described in Example 2. Singly protonated product withexact m/z and expected elemental formula matching the singly protonatedform of N-[2-(5-amino-7-methyl-1H-indol-3-yl)ethyl]acetamide havingchemical formula (XVI):

eluted at 2.3 minutes (EIC, see: FIG. 18A). Although peak splitting canbe minimized through the use of DMSO as injection solvent (Kaufman andJegle 2005, Agilent Technologies Technical Bulletin 5989-2485EN), thisphenomenon persisted owing to ion pairing effects between matrixcomponents (Tarafder et al. 2010, J Chromatogr A 1217:7065-7073).

As per standard procedures (Menéndez-Perdomo et al. 2021, Mass Spectrom56: 34683) high energy collisions (HCD) were achieved in a dedicated,post-LTQ, nitrogen collision cell. Orbitrap-based, HR fragment detectionwas employed (normalized collision energy, NCE 35), enabling opportunityto assign elemental formulae to subsequent diagnostic ion speciescharacteristic of a compound of formula (XVI), as follows (FIG. 18B,Table VII) (Servillo L. et al., 2013, J. Agric. Chem. 61: 5156-5162).

TABLE VII % Relative Ionic m/z Abundance Fragment Δ ppm 173.10714 100.00[M + H − C₂H₅NO]⁺ 1.04 232.14449 11.17 [M + H]⁺ 0.26 215.11788 9.63117.99882 0.95 201.97239 0.95 214.13376 0.63

Example 10—Biochemical Synthesis of a Twelfth Aminated PsilocybinDerivative

Escherichia coli strain Ec-3 was used to biosynthesize aminatedpsilocybin derivative, where the amino group terminating the 2-carbonaliphatic side chain was singly methylated. Ec-3 was constructed usingthe same method as for Ec-1 (see: Example 2), except that an additionalplasmid was assembled and transformed into cells along withpETM6-H10-TmTrpB-2F3-V5-BaTDC-FLAG. This additional plasmid encoded apromiscuous and efficient Rhinella marina N-methyltransferase enzymenamed RmNMT (SEQ. ID NO: 14). This additional plasmid was assembled asfollows: from plasmid pCDM4 (SEQ. ID NO: 1), the plasmidpCDM4-RmNMT-FLAG was created by inserting an in-frame, C-terminallyFLAG-tagged (SEQ. ID NO: 2, SEQ. ID NO: 3) RmNMT gene (SEQ. ID NO: 13)into the NdeI/XhoI site of pCDM4. The two target plasmidspCDM4-RmNMT-FLAG and pETM6-H₁₀-TmTrpB-2F3-V5-BaTDC-FLAG were transformedinto BL21 (DE3) cells, and antibiotics ampicillin plus streptomycin wereused to select for the correct clones containing both plasmids. Scaledup culturing and analysis were performed as described in Example 2,except that 6-methyl-1H-indol-4-ylamine (BLDPharm, www.bldpharm.com) wasused in place of 5,7-dimethyl-1H-indol-4-ylamine. To assess product,high-resolution LC-HESI-LTQ-Orbitrap-XL MS analysis was conducted asdescribed in Example 2. Singly protonated product with exact m/z andexpected elemental formula matching the singly protonated form of6-methyl-3-[2-(methylamino)ethyl]-1H-indol-4-amine having chemicalformula (XVII):

eluted at 2.4 minutes (EIC, see: FIG. 19 ). Although peak splitting andpoor peak shape can be minimized through the use of DMSO as injectionsolvent (Kaufman and Jegle 2005, Agilent Technologies Technical Bulletin5989-2485EN), this phenomenon persisted owing to ion pairing effectsbetween matrix components (Tarafder et al. 2010, J Chromatogr A1217:7065-7073).

1. A chemical compound or salt thereof having formula (I):

wherein at least one of R₄, R₅, R₆, or R₇ is an amino group or anN-substituted amino group, and wherein each non-aminated R₅, R₆, or R₇is a hydrogen atom, an alkyl group, or O-alkyl group, wherein R₂ is ahydrogen atom, wherein R₄ when it is not aminated is a hydrogen atom, anO-alkyl group, or a hydroxy group, and wherein R_(3a) and R_(3b) areeach independently a hydrogen atom, an alkyl group, an aryl group, or anacyl group.
 2. A chemical compound or salt thereof according to claim 1,wherein R₅ is an amino group or N-substituted amino group, and R₆ and R₇each are a hydrogen atom.
 3. A chemical compound or salt thereofaccording to claim 1, wherein R₅ is an amino group, and R₆ and R₇ eachare a hydrogen atom.
 4. A chemical compound or salt thereof according toclaim 1, wherein R₅ is an amino group, and R₆ and R₇ each are a hydrogenatom, and R₄ is an O-alkyl group.
 5. A chemical compound or salt thereofaccording to claim 1, wherein R₅ is an amino group, and R₆ and R₇ eachare a hydrogen atom, and R₄ is a (C₁-C₆)—O-alkyl group.
 6. A chemicalcompound or salt thereof according to claim 1, wherein R₅ is an aminogroup, and R₆ and R₇ each are a hydrogen atom, and R₄ is a(C₁-C₃)—O-alkyl group.
 7. A chemical compound or salt thereof accordingto claim 1, wherein R₅ is an amino group, and R₆ and R₇ each are ahydrogen atom, and R₄ is a methoxy group (—C(═O)CH₃).
 8. A chemicalcompound or salt thereof according to claim 4, wherein R_(3a) and R_(3b)are each independently a hydrogen atom or an acyl group.
 9. A chemicalcompound or salt thereof according to claim 4, wherein R_(3a) and R_(3b)are each a hydrogen atom.
 10. A chemical compound or salt thereofaccording to claim 1, wherein R₇ is an amino group or N-substitutedamino group, and R₅ and R₆ each are a hydrogen atom.
 11. A chemicalcompound or salt thereof according to claim 1, wherein R₇ is an aminogroup, and R₅ and R₆ each are a hydrogen atom.
 12. A chemical compoundor salt thereof according to claim 1, wherein R₇ is an amino group, andR₅ and R₆ each are a hydrogen atom, and R₄ is an O-alkyl group.
 13. Achemical compound or salt thereof according to claim 1, wherein R₇ is anamino group, and R₅ and R₆ each are a hydrogen atom, and R₄ is a(C₁-C₆)—O-alkyl group.
 14. A chemical compound or salt thereof accordingto claim 1, wherein R₇ is an amino group, and R₅ and R₆ each are ahydrogen atom, and R₄ is a (C₁-C₃)—O-alkyl group.
 15. A chemicalcompound or salt thereof according to claim 1, wherein R₇ is an aminogroup, and R₅ and R₆ each are a hydrogen atom, and R₄ is a methoxy group(—OCH₃).
 16. A chemical compound or salt thereof according to claim 12,wherein R_(3a) and R_(3b) are each independently a hydrogen atom or anacyl group.
 17. A chemical compound or salt thereof according to claim12, wherein R_(3a) and R_(3b) are each a hydrogen atom.
 18. A chemicalcompound or salt thereof according to claim 1, wherein R₇ is asubstituted amino group, and R₅ and R₆ each are a hydrogen atom.
 19. Achemical compound or salt thereof according to claim 1, wherein R₇ is anN-substituted amino group.
 20. A chemical compound or salt thereofaccording to claim 1, wherein R₇ is an N-substituted amino group,wherein the N-substituted group has the formula:

wherein R′ and R″ are each independently selected from a hydrogen atom,an alkyl group, and an aryl group.
 21. A chemical compound or saltthereof according to claim 20, wherein R′ and R″ are each independentlyselected from a hydrogen atom, and an alkyl group.
 22. A chemicalcompound or salt thereof according to claim 20, wherein R′ is a hydrogenatom and R″ is an alkyl group.
 23. A chemical compound or salt thereofaccording to claim 20, wherein R′ is a hydrogen atom and R″ is a(C₁-C₃)-alkyl group.
 24. A chemical compound or salt thereof accordingto claim 1, wherein R₇ is a substituted amino group, and R₄, R₅ and R₆each are a hydrogen atom.
 25. A chemical compound or salt thereofaccording to claim 24, wherein R_(3a) and R_(3b) are each independentlya hydrogen atom or an acyl group.
 26. A chemical compound or saltthereof according to claim 24, wherein R_(3a) and R_(3b) are each ahydrogen atom.
 27. A chemical compound or salt thereof according toclaim 1, wherein the chemical compound is selected from the groupconsisting of compounds having formulas (III); (IV); (V); (VI); (VII);(VIII); (IX); (X); (XI); (XII); (XIII); (XIV); (XV); (XVI); and (XVII):


28. A chemical compound or salt thereof according to claim 1, whereinthe chemical compound is at least about 95% (w/w) pure.
 29. Apharmaceutical or recreational drug formulation comprising an effectiveamount of a chemical compound or salt thereof according to claim 1,together with a pharmaceutically acceptable excipient, diluent, orcarrier.
 30. A method for treating a psychiatric disorder, the methodcomprising administering to a subject in need thereof a pharmaceuticalformulation comprising a chemical compound or salt thereof according toclaim 1, wherein the pharmaceutical formulation is administered in aneffective amount to treat the psychiatric disorder in the subject.